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Optical Sensors for
Biomedical Diagnostics
and Environmental
Monitoring
Optical Sensors for
Biomedical Diagnostics
and Environmental
Monitoring
Banshi Dhar Gupta

Anand Mohan Shrivastav
Sruthi Prasood Usha
CRC Press
Taylor & Francis Group
6000 Broken Sound Parkway NW, Suite 300
Boca Raton, FL 33487-2742
© 2018 by Taylor & Francis Group, LLC

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Library of Congress Cataloging-in-Publication Data
Names: Gupta, Banshi Dhar, author. | Shrivastav, Anand Mohan, author. | Usha,
Sruthi Prasood, author.
Title: Optical sensors for biomedical diagnostics and environmental
monitoring / Banshi Dhar Gupta, Anand Mohan Shrivastav, Sruthi Prasood Usha.
Description: Boca Raton, FL : CRC Press, Taylor & Francis Group, [2017] |
Includes bibliographical references and index.
Identifiers: LCCN 2017023196| ISBN 9781498789066 (hardcover ; alk. paper) |
ISBN 1498789064 (hardcover ; alk. paper) | ISBN 9781315156033 (eBook) |
ISBN 1315156032 (eBook) | ISBN 9781498789073 (eBook) | ISBN 1498789072
(eBook) | ISBN 9781351652100 (eBook) | ISBN 1351652109 (eBook) | ISBN
9781351642569 (eBook) | ISBN 1351642561 (eBook)
Subjects: LCSH: Optical fiber detectors. | Environmental monitoring. |
Plasmons (Physics)
Classification: LCC TA1815 .G87 2017 | DDC 681/.25--dc23
LC record available at https://lccn.loc.gov/2017023196
Visit the Taylor & Francis Web site at

http://www.taylorandfrancis.com
and the CRC Press Web site at
http://www.crcpress.com
Contents
Preface.......................................................................................................................ix
Acknowledgments..................................................................................................xi
About the Authors............................................................................................... xiii
1 Introduction to Sensors..................................................................................1
1.1 What Is a Sensor?................................................................................... 1
1.2 Need of Sensors......................................................................................3
1.2.1 Biosensors..................................................................................4
1.2.2 Gas Sensors................................................................................5
1.2.3 Chemical Sensors......................................................................6
1.3 Sensor Components............................................................................... 7
1.3.1 Analyte....................................................................................... 8
1.3.2 Recognition Unit....................................................................... 9
1.3.3 Transducer.................................................................................9
1.3.4 Detector/Analyzer.................................................................. 10
1.3.4.1 Spectrometer............................................................ 10
1.3.4.2 Optical Power Meter............................................... 10
1.3.4.3 Charge-Coupled Device......................................... 11
1.4 Sensor Performance Parameters........................................................ 11
1.5 Biosensor Classifications..................................................................... 12
1.5.1 Classification Based on Transducer..................................... 13
1.5.1.1 Electrochemical Transducer.................................. 14
1.5.1.2 Optical Transducer.................................................. 16
1.5.1.3 Mass-Sensitive Transducer.................................... 20
1.5.1.4 Calorimetric Transducer........................................ 20
1.5.1.5 Light-Addressable Potentiometric Transducer......21
1.5.2 Classification Based on Bio-Receptors................................. 21
1.6 Biosensor Regeneration....................................................................... 27
1.7 Overview of the Book.......................................................................... 29
References........................................................................................................ 30
2 Basics of Resonance...................................................................................... 33
2.1 Resonance-Based Sensors................................................................... 33
2.2 SPR for Sensing....................................................................................34
2.2.1 Theory of SPR..........................................................................34
2.2.2 Theory of LSPR........................................................................44
2.2.3 Realization of Sensors............................................................ 48
2.3 Types of Resonances............................................................................ 55
2.3.1 Surface Plasmon Resonance.................................................. 55
v
vi Contents
2.3.1.1 Long-Range SPR...................................................... 55

2.3.1.2 Short-Range SPR...................................................... 58
2.3.1.3 Nearly Guided Wave SPR....................................... 58
2.3.1.4 Waveguide-Coupled SPR....................................... 59
2.3.1.5 Magneto-Optic SPR................................................ 59
2.3.1.6 Fano Resonance SPR............................................... 60
2.3.1.7 Gap SPR.................................................................... 61
2.3.2 Lossy Mode Resonance.......................................................... 61
2.3.3 Interferometric Resonance..................................................... 61
2.3.3.1 Fabry–Perot Interferometric Resonance............... 62
2.3.3.2 Michelson Interferometric Resonance.................64
2.3.4 Acoustic Resonance................................................................64
2.4 Developing LMR for Sensing Applications......................................64
2.5 Summary............................................................................................... 70
References........................................................................................................ 72
3 Fiber-Optic Sensors...................................................................................... 75
3.1 Optical Fiber as Sensor Element........................................................ 75
3.2 Factors Affecting Light Propagation.................................................77
3.2.1 Numerical Aperture and Acceptance Angle...................... 78
3.2.2 V-Number and Fiber Modes..................................................80
3.2.3 Fiber Parameters..................................................................... 81
3.2.4 Evanescent Wave..................................................................... 82
3.3 Advantages of Fiber-Optic Sensors...................................................84
3.4 Parameters Tailoring Sensor Performance....................................... 85
3.4.1 Sensitivity................................................................................. 85
3.4.2 Selectivity................................................................................. 87
3.4.3 Limit of Detection................................................................... 88
3.4.4 Limit of Quantification.......................................................... 89
3.4.5 Repeatability............................................................................ 89
3.4.6 Reproducibility........................................................................ 90
3.4.7 Detection Accuracy................................................................90
3.4.8 Figure of Merit.........................................................................90
3.5 Designs of a Fiber-Optic Sensor Probe............................................. 91
3.5.1 Straight Probe.......................................................................... 91
3.5.2 U-Shaped Probe...................................................................... 94
3.5.3 Tapered Probe.......................................................................... 97
3.5.4 D-Shaped Probe...................................................................... 99
3.6 Summary............................................................................................. 100
References...................................................................................................... 100
4 Nanostructured Sensors............................................................................. 103

4.1 Nanotechnology as a Sensing Platform.......................................... 104
4.2 Metallic Nanostructures and Synthesis.......................................... 107
4.2.1 Silver....................................................................................... 109
Contents vii
4.2.1.1 Chemical Methods................................................ 109

4.2.1.2 Physical Methods.................................................. 112
4.2.1.3 Biological Methods................................................ 118
4.2.2 Gold......................................................................................... 119
4.2.3 Platinum................................................................................. 123
4.2.4 Palladium............................................................................... 126
4.3 Effect of Nanostructures................................................................... 127
4.3.1 Shape of Nanoparticles........................................................ 129
4.3.1.1 Spherical................................................................. 129
4.3.1.2 Others...................................................................... 130
4.3.1.3 Magnetic Nanoparticles....................................... 130
4.3.2 Nanorods................................................................................ 131
4.3.3 Nanowires.............................................................................. 131
4.3.4 Other Nanostructures.......................................................... 132
4.4 Nanostructures for Sensing.............................................................. 133
4.4.1 Single-Nanoparticle Sensors............................................... 135
4.4.2 Surface-Enhanced Nanosensors......................................... 137
4.5 Applications........................................................................................ 140
4.5.1 Detection of Physical Parameters....................................... 140
4.5.2 Environmental and Agricultural Monitoring.................. 141
4.5.3 Biological Applications and Biomarkers........................... 150
4.5.4 Surgical and Clinical Diagnostics...................................... 155
4.6 Summary............................................................................................. 157
References...................................................................................................... 157
5 Semiconductor Metal Oxide Sensors...................................................... 165

5.1 Role of SMO in Sensor Applications............................................... 166
5.2 Properties Supporting Sensing........................................................ 170
5.2.1 Surface and Structure........................................................... 170
5.2.2 Conductivity.......................................................................... 172
5.2.3 Catalytic/Chemical Activity and Stability........................ 172
5.2.4 Sensitivity and Reversibility............................................... 173
5.3 Nanostructured Metal Oxides......................................................... 174
5.4 Mechanism of Gas Sensing and Applications............................... 176
5.4.1 SPR-Based SMO Gas Sensors.............................................. 178
5.4.2 LMR-Based SMO Gas Sensors............................................ 183
viii Contents
5.5 Biosensing with SMOs...................................................................... 189

5.6 Summary............................................................................................. 194
References...................................................................................................... 195
6 Molecular-Imprinting-Based Sensors..................................................... 197

6.1 Basics of Molecular Imprinting........................................................ 198
6.1.1 Molecular-Imprinting Elements......................................... 199
6.1.2 Synthesis Protocols............................................................... 204
6.2 Types of Molecular Imprinting........................................................ 207
6.2.1 Covalent Molecular Imprinting.......................................... 207
6.2.2 Noncovalent Molecular Imprinting................................... 208
6.2.3 Molecular Imprinting in Nanostructures......................... 209
6.3 Molecular-Imprinting Polymer as a Floor for Sensing................. 211
6.4 Applications........................................................................................ 212
6.4.1 Food Safety............................................................................ 212
6.4.2 Environmental Monitoring................................................. 217
6.4.3 Biomedical Uses.................................................................... 219
6.5 Summary.............................................................................................222
References......................................................................................................223
7 Summary and Future Outlook.................................................................. 227

Index...................................................................................................................... 231
Preface
This book is an effort to bring a comprehensive book on optical sensors for

biomedical diagnostics and environmental monitoring for the undergradu-
ate and graduate students apart from researchers working in various related
fields. The idea of writing this book came during a group discussion where
it was felt that a detailed information on various kinds of sensors realized on
this topic incorporating the newly implemented and upcoming techniques
should be provided to the community because of their direct relation to the
health of the living body like human beings, animals, etc. Various kinds of
newly developed techniques are being used for the development of sensors
for such applications. Surface plasmon resonance (SPR) is one of such tech-
niques which has been widely used and can be easily understood with the
elementary knowledge of electromagnetic theory and hence can be recom-
mended to the students with its possible applications. The literature has also
shown the upcoming technique of lossy mode resonance (LMR) for sens-
ing applications. LMR exhibited by the semiconductor metal oxide (SMO) in
the visible region and possessing applications in sensing of various gases,
chemical and biological analytes also need wide attention. The invention of
optical fiber technology has made easy implementation of SPR- and LMR-
based sensors with the unique advantages of flexible designs with capabil-
ity of online monitoring and remote sensing using cost-effective substrate.
Nanotechnology is an intense branch that has made its role clear in sensing
applications by its irreplaceable performances in many aspects. However,
the integration of optical fiber and nanotechnology has fascinated the world
of the sensors. Therefore, a comprehensive collective knowledge of this inte-
gration and its advantages in the development of sensors is required. The
very new field of molecular imprinting is gaining its attraction mainly in the
field of biomolecular recognition. This book is the first work to explain in
detail the usage of this method mainly in fiber-optic sensors and, in particu-
lar, in integrating it with the nanoparticles for recognizing biosensors. With
this book, we aim to introduce the graduate students and early research-
ers to an overview of the plasmonic and LMR-based sensors for biomedical
diagnostics and environmental monitoring employing the miracles of the
fascinating fields, their working principles, and applications.
This book will benefit the researchers as well as the industrial and pub-
lic society with an overview of recent advances of optical sensors and their
applications for human health and safety. This book starts with the introduc-
tion of sensors, need of sensors, and distinct types of sensors applicatory for
biomedical and environmental monitoring (Chapter 1). Biomedical applica-
tions of sensors are gaining importance intently and hence a deep attention
is given to biosensors for the diagnosis of various diseases important for
ix
x Preface
healthcare in this chapter. Chapter 2 introduces the basic concepts of reso-

nance and various resonance-based sensing techniques that are currently
being used in various research and industrial applications. SPR and its
collateral techniques are discussed in detail along with the chronological
development of the field. In addition, a new technique, LMR, developing its
application in the sensing, is also explored. In Chapter 3, a discussion about
the fiber-optic sensors, the essential components of fiber-optic sensors, their
functions, and the parameters tailoring sensor performance are presented.
The multiple design possibilities such as D-shaped, tapered and U-shaped
miniaturized probe, in addition to the uniform straight core probe, are
included. This is followed by the discussion on nanostructures and their
role as a sensing platform in Chapter 4. The chapter includes various metal-
lic nanostructures useful in sensing and their fabrication methods. A num-
ber of sensors utilizing nanostructures are presented for environmental and
agricultural monitoring, biological applications, biomarkers, surgical, and
clinical diagnostics. In Chapter 5, an analysis on the action of SMO for sens-
ing purpose is presented. The section illustrates the SPR and LMR mecha-
nism of sensing, involving metal oxide, for the monitoring of gases, their
upcoming applications in biomolecular detection and diagnosis. Chapter 6
introduces the latest technique of molecular imprinting incorporating the
qualitative analysis and amelioration done by the technique in the field of
sensing applications. The sensors realized using molecular imprinting for
food safety, environmental monitoring, and biomedical usage are discussed
in the chapter. The last chapter of this book (Chapter 7) summarizes all the
contents with a note on future trends of research in the field of fiber-optic
and other technique-based sensors for environmental monitoring and bio-
molecular recognition. It is hoped that this book will serve the purpose for
which it has been written.
Banshi Dhar Gupta

Anand Mohan Shrivastav
Sruthi Prasood Usha
Physics Department
Indian Institute of Technology Delhi
New Delhi, India
Acknowledgments
I consider myself very fortunate to have worked with Dr. Chandra Deep
Singh, Professor Sunil Khijwania, and Dr. Navneet K. Sharma on fiber-optic
evanescent field absorption sensors. My continuous interaction with them
has made me wiser in the field of fiber-optic sensors. It was this interaction
that resulted in the publication of my first book titled Fiber Optic Sensors:
Principles and Applications (2006). In 2004, I took an initiative to work on fiber-
optic sensors utilizing surface plasmon resonance technique. The research
carried out on this topic resulted in 10 PhD theses. I thank Drs. Anuj K.
Sharma, Rajan Jha, Rajneesh K. Verma, Sachin K. Srivastava, Sarika Singh,
Priya Bhatia, Roli Verma, Satyendra K. Mishra, and Rana Tabassum who
have worked with me in this exciting field. A part of their work resulted in
my second book titled Fiber Optic Sensors Based on Plasmonics (2015). For the
last 5 years, my focus has been on the research and development of opti-
cal sensors for biomedical diagnostics and environmental monitoring. My
present graduate students, apart from Anand Mohan Shrivastav and Sruthi
Prasood Usha (also the authors of this book), Ravi Kant, Vivek Semwal,
Anisha Pathak, and Sonika Sharma are working in this area. I learnt a lot
from these colleagues through interaction on this topic. Indeed, a part of the
book has emerged from the work carried out with these graduate students
and with some of my old students. I thank all of them for their hard work
in the laboratory without which publication of this book was not possible.
Finally, special thanks to my wife Uma Gupta for her patience and under-
standing during the writing of this book.
Banshi Dhar Gupta
xi
About the Authors
Banshi Dhar Gupta earned his MSc in physics in 1975 from Aligarh Muslim
University, India and a PhD in physics in 1979 from the Indian Institute of
Technology, New Delhi. In 1978 he joined the Indian Institute of Technology,
New Delhi, where he is currently a professor of physics. In addition,
Professor Gupta has worked at the University of Guelph, Canada between
1982 and 1983, the University of Toronto, Canada in 1985, the Florida State
University, Florida, USA in 1988, the University of Strathclyde, UK in 1993,
and the University of Birmingham, UK in 2010. In 1992, he was awarded
the ICTP Associateship by the International Centre for Theoretical Physics,
Trieste, Italy, which he held for 8 consecutive years. In this capacity, he visited
ICTP, Italy in 1994 and 1996. Professor Gupta is a recipient of the 1991 Gowri
Memorial Award of the Institution of Electronics and Telecommunication
Engineers, India. He has published more than 170 research papers in interna-
tional journals and 100 papers in conferences. Professor Gupta has authored
books titled Fiber Optic Sensors: Principles and Applications (New India
Publishing Agency—NIPA, New Delhi, 2006) and Fiber Optic Sensors based
on Plasmonics (World Scientific, 2015), and is the coeditor of the Proceedings of
SPIE, USA, Vol. 3666, 1998 and Vol. 8173, 2010. Professor Gupta has delivered
invited talks on plasmonics-based sensors in various international confer-
ences held in the United States, China, Australia, Korea, and India. Professor
Gupta is a member of the editorial board of the Journal of Sensors. His current
areas of interest are plasmonic biosensors, fiber optic sensors, and nanotech-
nology. He is a regular member of the Optical Society of America and life
member of the Optical Society of India.
Anand Mohan Shrivastav earned his MSc in physics in 2012 from C.S.J.M.
University Kanpur, India. He joined IIT Delhi as a full time PhD student in
2013. Mr Shrivastav has published more than 15 research papers in interna-
tional journals and 10 papers in conferences. His current areas of interest
are fiber optic sensors, nanotechnology, and molecular imprinting technol-
ogy. Mr Shrivastav was awarded senior research fellowship in 2015 by the
University Grants Commission (UGC), India. He is a member of the Optical
Society of America and Optical Society of India.
Sruthi Prasood Usha earned her BTech in electronics and communication

engineering in 2010 from PAACET (P.A. Aziz College of Engineering and
Technology), Kerala University, India and MTech in optoelectronics and
laser technology in 2013 from Cochin University of Science and Technology
(CUSAT, India). Ms. Usha joined IIT Delhi as a full time PhD student
xiii
xiv About the Authors
in 2014. She has published 12 research papers in international journals

and 10 papers in conferences. Her current areas of research are fiber optic
SPR (surface plasmon resonance) and LMR (lossy mode resonance) sen-
sors, nanostructures, and nanotechnology. She is a member of the Optical
Society (OSA).
1
Introduction to Sensors
Measurements of various physical, chemical, and biological parameters/

analytes such as pressure, temperature, magnetic field, pH, pesticides, gases,
glucose, urea, cancer, etc. are required for various applications such as envi-
ronment and health monitoring, process control, and electrical utilities/
power plants. The devices used to measure these parameters are called sen-
sors. Tremendous amount of work has been carried out on the development
of low-cost, compact, sensitive, and analyte-specific sensors. Their progres-
sion over the decades has reached well-established pace due to the rapidly
developing technology and science. The sensors are named based on the
technique used by them for the measurement of abovementioned param-
eters. The technique can be optical, electrical, mechanical, electrochemical,
or any other. The present book deals with the biomedical diagnostics and
environmental monitoring sensors utilizing optical techniques. Below, we
shall first define a sensor and then discuss the important features of a sen-
sor such as need of a sensor, its components, performance parameters, and
classifications.
1.1 What Is a Sensor?

Sensor is a device which detects and analyzes a parameter continuously
by providing an output in a way that is readable by humans or human-
controllable instruments in a continuous manner with possibilities of repro-
ducibility and recovery. Further, in a sensor, change in one form of input
energy to corresponding single-valued change in another or same form of
energy takes place. The output of a sensor is usually electrical or optical in
nature. The sensors of the current decade can quantify and qualify almost all
the physical, mechanical, chemical, and biological entities. There are many
things which are expected from a sensor. First, the sensor should be insensi-
tive to the change in external environmental parameters such as tempera-
ture and humidity (for nontemperature and humidity sensors) to provide
the invariable results. Second, the allocation of a sensor to analyze a particu-
lar parameter in a natural environment should be made considering its abil-
ity to withstand harsh atmosphere conditions and changes, the domain in
which the sensor holds application, its lifetime, remote activities, and online
1
2 Optical Sensors for Biomedical Diagnostics and Environmental Monitoring
compatibility with good recovery. Considering all these aspects, the work
reported in the literature so far has concluded that the optical way of com-
mutating changes is better than the electrochemical and calorimetric means.
In optical sensors, the method of converting one form of energy to another
is optical; however, the system may make use of other fields such as electri-
cal, electronic, mechanical, biological, nanostructural, material, biochemical,
chemical, and physical engineering along with optics. Thus, the versatile-
integrative behavior of optical sensors covers a broad area of requirements.
The optical-sensing system, as in the case of all other methods, has four
basic units. The foremost unit is the light source of the sensor that needs to
be selected according to the design of the sensor system and the application
to which it is oriented. The second unit is the sensor receptor/recognition
element which interacts with the analyte (to be sensed) and changes its own
optical properties such as refractive index. The change in the properties of
recognition unit is determined by the third unit called as transducer. The
transducer is the unit which converts the change in the recognition unit prop-
erties into some measurable signal by means of an optical transducing phe-
nomenon. The measurable parameter of the signal can be the intensity, phase,
or wavelength of the light source which is detected with the help of detector/
analyzer which is the fourth and last unit of a sensor. Figure 1.1 shows the
schematic of a generic sensor with all the required units. The working of the
whole system depends on the mechanism as well as the principle on which
the sensor works and also on its utilization. For a sensing system based on
optical method, the variation in one of the properties of light such as ampli-
tude/intensity, phase, polarization, frequency, and wavelength due to the
change in the sensing medium is analyzed.
Optical-sensing system may use lens, prism, mirror, grating, microscope
objective, waveguide, fiber, etc. and optical detection as well as analyz-
ing units. The detection in optical system can be performed by schemes
of luminescence, fluorescence, and absorption. The advantages of optical
sensing include immunity to electromagnetic interference, possibilities of
point and distributed arrangement with multiplexing, compatibility, and
Analyte
Recognition
unit
Transducer Detector/
Source
Input unit Output analyzer
FIGURE 1.1
Schematic of a generic sensor with the essential units.
Introduction to Sensors 3
electrical isolation (Grattan and Sun 2000). The usage of fiber as the opti-
cal substrate competes with all other schemes of optical sensing, as well as
sensing based on transducer elements other than optical due to its capabil-
ity of remote sensing and online monitoring. Hence, in this book, we shall
impart the significance of optical sensing, mainly using optical fiber, for the
physical, chemical, and biological sensing to cover environmental supervi-
sion and biomedical diagnostic applications, which cover the relevant areas
of sensor operation in present and future. The upcoming sections discuss
the relevance of sensors in detecting various biological, gaseous–chemical
measurands with the parameters which quantify the sensor performance.
Section 1.5 will discuss the types of classifications of sensors followed by
sensor regeneration aspects in Section 1.6. Finally, Section 1.7 contests the
framework of the book.
1.2 Need of Sensors

The sensors are one of the most important devices that we use in day-to-day
life. As we all know, our nervous system consists of five vital sensing organs:
nose, skin, eyes, ears, and tongue. These are used for the sensing of olfac-
ception (smell), tactioception (touch), ophthalmoception (sight), audioception
(listening), and gustaoception (taste), respectively. Various types of devices
and machines are equipped with sensors that help them in working. Consider
a mobile phone, it is a device combined with multisensor technology such as
accelerometer and gyroscope that are used for the determination of linear
and angular rotations. Various types of temperature and humidity sensors
are being used daily in several electronic machines such as refrigerators,
air conditioner, and air cooler for the automated working. Light-dependent
resistors (LDRs) are used as sensors in the automatic doors and streetlights
with an ability to switch their operation. Apart from these, sensors are also
used in various fields for the human and environmental safety, biomedi-
cal applications, military applications, etc. The aim of sensor development
for the environmental monitoring is the safeguard of the living organisms
from a number of pathogens and toxic contaminants that are continuously
released into the environment, resulting in the pollution of air, soil, and water
(Rodriguez-Mozaz et al. 2004). Sensors are also required in various biomedi-
cal and clinical applications to detect glucose, urea, vitamins, and nutrients
concentrations in human body, and also need to be detected for ensuring
their level within the normal range in the body. In order to ensure the safety
of food and water, several types of sensors for the detection of impurities
such as heavy metal ions (to secure for the use of hard drinking water), pes-
ticides, melamine, etc. are used and further advancements in these sensors
are required for highly specific and selective detection. In accordance with
the applications of sensors for biomedical, clinical, environment, and food

safety, the sensors can be classified in three broad categories as biosensors,
gas sensors, and chemical sensors.
1.2.1 Biosensors
Biosensor is a device which is generally designed and fabricated with a bio-
logical sensing element incorporating a transducer. It generates a change in
the transducer signal or bio-recognition element characteristics on exposure
to an analyte for which it has been designed. The change in signal depends
upon the concentration of the analyte. A number of definitions have been
given to the biosensor by the people working in the field of biosensing. The
general definition is: “Biosensor is a device that utilizes biological compo-
nents such as enzymes, antigens/antibodies for the determination of the con-
centration of the biomaterials” (Wang and Wolfbeis 2013). The biomaterials
to be detected are usually the materials which are toxic in nature and can
harm human body as well as the environment. The invention of first biosen-
sor was by Leland Clark and hence he was renowned as the father of the
biosensing concept (Clark and Lyons 1962). He showed detection of glucose
using oxygen electrode, where glucose oxidase was entrapped over a Clark
oxygen electrode using dialysis membrane. The estimation of glucose con-
centration was performed by measuring the reduction in the dissolved oxy-
gen concentration. The idea proposed was industrialized by Yellow Springs
Instrument Company (Ohio, USA) in 1975 and the glucose sensing analyzer,
based on the amperometric detection method, was successfully relaunched.
Later, the detection of urea concentration using direct potentiometric method
was reported (Katz and Rechnitz 1963). Further development in the sensor
was made with the introduction of urea electrode (Guilbault and Montalvo
Jr 1969). The biosensor based on microbe technology was pioneered in 1976
(Cammann 1977) and the term “biosensor” was introduced. The technol-
ogy was further developed with the application of artificial redox mediators
(Kulys and Svirmickas 1980) and later, the first ferrocene-mediated ampero-
metric glucose biosensor was proposed (Cass et al. 1984), which was indus-
trialized by Medi Sense Inc. in 1987. As a continuation of the work done by
various researchers, IUPAC introduced the definition of biosensors analogous
to chemosensors as “Biosensor is a device that uses specific biochemical reac-
tions mediated by isolated enzymes, immune-systems, tissues, organelles or
whole cells to detect chemical compounds usually by electrical, thermal or
optical signals” (Monosik et al. 2012). Followed by this great achievement,
a number of research communities from various fields like very-large-scale
integration (VLSI), life science, physics, chemistry, and material science initi-
ated their interest for the development of biosensing devices incorporating
more sophisticated, reliable, and miniaturized biosensing capabilities. In
2007, an implanted glucose biosensor (freestyle navigator system) was oper-
ated for 5 days and the work on the development of biosensors continued its
advancement exploiting the enzymes, antibodies, and microbes in combina-

tion with various types of transducers (Weinstein et al. 2007).
The term “biosensor” signifies a device which is fabricated by the combi-
nation of two parts: bio-recognition element and the sensor element. In the
biosensors, bio-recognition elements like enzyme and antigen recognize the
selective analyte to be detected, while the sensor element is used for the trans-
duction of the change in biomolecule concentration into an electrical/optical
signal. Hence, the sensor element is also recognized as the transducer. The
bio-recognition element should be highly specific to the analyte, to which it
is sensitive for the change in the electrical/optical signal. Biosensors are used
for industrial applications, biomedical diagnostics, food safety, and military
applications. In the beginning, major application of biosensors was the sens-
ing of glucose concentration in blood due to its plentiful application in bio-
medical industry. However, nowadays, biosensors are used in a number of
commercialized applications such as fabrication of devices for the calibration
of foods and beverages concerning the human health, development of the
devices for environmental sampling, and noninvasive instruments for clini-
cal analysis (Mello and Kubota 2002, Patel 2002). However, the industrializa-
tion of biosensor devices is quite slow due to various technological issues
and limitations. For example, it is necessary to prevent the biosensor from
contamination and is challenging too at the same time, because of the pres-
ence of semiconductor materials along with the biomolecules in biosensing
devices.
1.2.2 Gas Sensors

The development of the gas sensors became the topic of discussion after the
discovery of the effect of harmful gases on human health. During the nine-
teenth and early twentieth centuries, coal miners used to take canaries to
the coal tunnels for the early detection of life-threatening gases such as car-
bon monoxide, carbon dioxide, and methane. Canary, a very songful bird,
stops singing and eventually dies in the continuous presence of gases. This
was used as a signal for the detection of gases (Thompson 2004). The first
gas detection and monitoring system was a flame safety lamp invented by
Darway in 1815. In the lamp, the height of the oil flame was adjusted to a spe-
cific position in fresh air which was kept inside the glass sleeve with a flame
arrester. High flame was the indicator of the presence of methane gas, while
the low flame was for low oxygen. Later, during 1926–1927, Oliver Johnson
introduced a gas sensor based on catalytic combustion method. After this,
various types of gas sensors based on methods such as electrochemical and
catalytic combustion were commercialized. In the 1970s, further establish-
ment of various types of gas sensors was carried out with different tech-
niques including humidity analysis, semiconductor combustibility, and
solid electrolyte oxygen. Later, extensive attempts were made not only for
the advancement of these sensors but also for the implementation of several
types of new gas sensors, which were in great demand for various applica-
tions such as environmental reservation, energy saving, health safety, air
pollution monitoring, etc. Gas sensors are the most important part of envi-
ronmental monitoring system because of the gases produced from various
sources such as home appliances (chimneys to air conditioners) and various
industries that expel toxic waste products. Hence, continuous monitoring of
environment is required for human safety and unpolluted atmosphere. Due
to these reasons, the gas-sensors-producing industries have been developed
up to a significant level to meet the requirements of providing a standard
and clean atmosphere for better living. However, further improvement in the
gas-sensing technology is required for the improvement of the quality of life
and sustainability of our society.
1.2.3 Chemical Sensors

Chemical sensor is used for the determination of chemical composition of
the surrounding medium which can be either in gaseous phase or in liquid
phase. If the target to be detected is in the gaseous phase, then the device
prepared for the detection of gases is generally termed as the gas sensor.
The sensor recognizes the presence of the target species and transforms it
into a physical signal. Thus, a variation in the characteristic physical signal
is observed due to the variation in the concentration of target species. Being
a broad area, the technique of gas sensing is now considered as a separate
field. The first significant work for the development of chemical sensor was
reported by Arnold Backman in 1932 with the invention of modern glass
electrode for sensing application, which was later commercialized. Later,
in 1936, he designed and patented a pH meter based on a pair of glass elec-
trodes. In the patented device, the electrode pair was suspended in the test
solution and the potential difference between the electrodes was used to
measure the pH value of the test solution. In 1937, the application of solid-
state electrodes such as silver halide and fluoride-specific electrodes for
chemical sensing was introduced (Kolthoff and Sanders 1937). The chemical
sensors utilizing antibodies as the recognition system for the detection of
biological samples were developed (Moore and Pressman 1964). Almost 2
years later, the selective and reversible properties of alkali metal ions were
reported (Stefanac and Simon 1966). This study was further used for the
development of K+-selective electrochemical sensor (Frant 1994). An elec-
trochemical serum analyzer for NASA’s Space Shuttle was proposed and
the first industrialized sodium/potassium analyzer in blood sample was
reported, which was named as SS-30.
Followed by the development of chemical sensors based on naturally
occurring antibodies with highly specific properties, the researchers
started focusing on the fabrication of artificial ligands for the detection
of various analytes. The studies for the development of artificial recep-
tors were pioneered by Pederson, Lehn, and Cram who were awarded the
Nobel Prize in 1987 (Pedersen 1967, Cram 1988, Lehn 1988). Various syn-
thetic ligands such as synthetic macrocyclic polyethers, crown compounds,
cyclophanes, and macrohetero-bicyclic were developed for the chemical
sensor applications by the team. In the same period, an optical chemi-
cal sensor was introduced by evaluating the first optical potassium test
based on dry reagent chemistry (Keller 2008). The method was commer-
cialized by Ames Division. In 1968, the first fluorosensor for oxygen detec-
tion was used for the biomedical applications (Lubbers and Optiz 1975).
Lubbers suggested the name of optical sensor as, “optrode” analogous to
the “electrode,” which was further corrected as “optode.” A considerable
amount of work was carried out by various research groups in 1980s using
optical chemical sensors for food safety and environmental monitoring.
Later in 1991, after the remarkable growth of the chemical sensor industry,
IUPAC gave the definition of the chemical sensor as, “A chemical sensor
is a device that transforms chemical information, ranging from concentra-
tion of specific sample component to total composition analysis, into an
analytically useful signal” (Hulanicki et al. 1991). Few more definitions
of the chemical sensors were reported in a more general way by various
authors. A well-known one by Wolfbeis is as, “Chemical sensors are small
sized devices comprising a recognition element, a transduction element
and a signal processor capable of continuously and reversibly reporting a
chemical concentration” (Wolfbeis 1991).
1.3 Sensor Components

As mentioned above, the sensor can be divided into four components: ana-
lyte, recognition unit, transducer, and analyzer (Diamond 1998). The work-
ing of the sensor is based on the interaction of analyte to be sensed with the
recognition unit, which makes a significant change in the characteristics of
the input signal that is detected with the help of a transducer, by a detec-
tion unit or analyzer. The analyte/sample may be biological (such as glucose,
cholesterol, proteins, etc.), chemical (such as pH, metal ions, etc.), and envi-
ronmental (gases like CO2, CH4, etc.). Recognition unit is the most important
component of the sensor. The analyte to be detected reacts with the recogni-
tion unit through a predesigned interaction. Numerous types of natural and
artificial receptors have been introduced by researchers for developing the
sensing devices. These will be discussed in further sections. Transducer is
the next salient component of the sensing unit. It is used for the conversion of
one type of signal into other form, thus converting the analyte concentration
into corresponding signal. Analyzer or detector is the last but essential part
of the sensing unit. The analyzer is required for detecting/measuring sev-
eral types of signals which correspond to recognition of the analyte. A block
Analytes:
Air pollutants, human
samples (blood, urine, and
Reaction saliva), pesticides, etc.
Receptors:
Enzymes, antibodies,
microbes, artificial receptors
Recognition
of the Immobilization
reaction
Transducers:
Optical fiber, metals, prism,
electrodes, nanostructures
Observation
Analyzer:
Voltmeter, spectrometer,
power meter, CCD
FIGURE 1.2
Block diagram of the sensor components, showing the analyte/s that needs to be interacted
with recognition element/s interfaced with the transducer and then the results of interaction
are transferred to the detection and analysis unit.
diagram showing the sensor components with possibilities in their selection

and path in which the working proceeds is shown in Figure 1.2.
1.3.1 Analyte
Analytes (samples) are various types of atmospheric, chemical, biological
elements which need to be detected. The analyte may be either in liquid state
or in gaseous form. According to the type of analyte, sensors can also be
classified as chemical, gas, or biosensors. Various hazardous gases, atmo-
spheric pollutants, chemical wastes through industries, biological elements
present in blood/urine, nutrients and vitamins, pesticides, etc. are the few
examples of analytes needed to be sensed. Several types of ambient condi-
tions and physical parameters such as pH, temperature, and pressure also
need to be sensed as they play important roles in the industries for the fabri-
cation of various medicines, devices and need to be monitored for the human
and environmental safety. Thus, these can also be considered as analytes.
Most common gases such as hydrogen, hydrogen sulfide, chlorine, methane,
carbon monoxide, water vapor (humidity), ethanol vapor, etc. can be treated
as analytes in the gas-sensing unit. The concentration of metal ions in water
should be optimized because higher concentration of various metals such
as lead, calcium, cadmium, iron, arsenic can result in unhealthy conditions.
The vitamins A, B, C, D, E, and K are essential for human body which can
be found in various food stuffs such as egg, milk, fish, etc. except vitamin D,
which is obtained from direct sunlight. Each vitamin has its own functional-
ity and its deficiency causes many health-related problems in human body.
Thus, their concentration/availability should be sensed in various foodstuffs
as well as in blood. In blood, the analysis of various biological elements for
clinical applications is possible. Other analytes such as pesticides, minerals,
antibodies, and antibiotics are required to be detected for a healthy life. The
concentration of analyte is recognized by its interaction with the receptor/
recognition unit.
1.3.2 Recognition Unit

Recognition unit, generally termed as the receptor, is the most important
part of any sensing device since it decides the interaction nature of the sen-
sor with the analyte and hence the result. This is also called as the “heart of
a sensor.” The recognition unit or receptor should be selective and specific,
while interacting with the target analyte to be sensed. It should interact with
the analyte reversibly, and specifically without any interference from other
similar chemical analytes except the target analyte. Generally, in sensors,
receptors are immobilized over the surface of the transducer, so that trans-
ducer can transform the interaction between analyte and recognition unit
into a readable output signal which can be analyzed by the analyzer. For
example, in the case of enzymatic biosensors, the enzyme which interacts
with a specific analyte is the receptor of the sensor. In the case of few physi-
cal sensors like temperature and pressure sensors, no receptor is required
as the working of the sensor is based on the physical properties of materials
used in the fabrication of the fiber/transducer. A detailed discussion on the
types of receptors and their compatibility with the transducer is given in the
next section.
1.3.3 Transducer
By definition, transducer is a device which converts one kind of signal into
another form. In the case of sensors, transducer is the unit which helps to
measure the change in the properties of the receptor due to its interaction
with the analyte, by converting the changes in to some readable signals. In
optical sensors, the readable signal is the change in the parameters of the
output light collected by a detector/analyzer. The phenomenon by which
this conversion is performed is the transducing phenomenon which can be
optical, electrical, etc. Further, the element which realizes the transducing
phenomenon is called as the transducer such as metal films, nanoparticles,
fluorophores, etc. Generally, the recognition unit is immobilized over the
transducer surface which is sensitive toward the changes over its surface
properties. When the analyte interacts with the receptor unit, the optical
properties of the receptor unit change. Since the receptor is immobilized over
the transducer surface, the properties of transducer surface change and due
to the transducing mechanism (such as surface plasmon resonance [SPR]),
the change in the specific parameter (such as minimum transmission at a
specific wavelength, resonance wavelength) is obtained at the detection unit.
Thus, the light from the source interacts with the optical transducer unit and
is collected at the output after undergoing changes in its parameters. The
change in the parameter value specifies the interaction and sensitiveness of
the transducer surface to the analyte.
1.3.4 Detector/Analyzer
After the working of the transducer, the change in the signal is measured by
the detector. These are selected according to the transducer used, for exam-
ple, thermometer for measuring the temperature, ammeter for the measure-
ment of current, voltmeter for the voltage observation, magnetometer for
analyzing the magnetic field, optical power meter for measuring the light
intensity, and so on (Gupta 2006). There are various types of optical detectors
such as spectrometer, optical power meter, photodetector, charge-coupled
device (CCD), fluorometer, etc. Since the present book deals with the opti-
cal sensors, we shall now provide a brief introduction of some of the optical
detectors used in the optical sensors.
1.3.4.1 Spectrometer
Spectrometer is one of the most broadly used detectors in optical sensors.
It is used for the measurement of light intensity for a particular wavelength
range. The spectrometer consists of an entrance slit, collimator, dispers-
ing element (prism or grating), focusing lens, exit slit, and a detector. The
grating-based spectrometer has resolution better than the prism-based spec-
trometer. The grating with few hundred lines per mm is placed at a specified
blaze angle, which decides the operating wavelength range of the spectrom-
eter. For the detector of the spectrometer, usually a CCD linear array with
thousands of pixels is used. These are responsible for the fast processing
of the data. The selection of all the components of a spectrometer such as
entrance slit, diffraction grating, order shorting filter, and the detector coat-
ing strongly affects the performance parameters of the spectrometer such as
spectral range, resolution, sensitivity, and stray light.
1.3.4.2 Optical Power Meter

Optical power meter is used for the measurement of the power in an opti-
cal signal. It is usually used for the determination of the power of the light
guided through fiber-optic cable. It is made up of a calibrated sensor inter-
faced with an amplifier and display. It is also used for the observation of
power loss in the optical signal when light is guided through the fiber. In
the case of sensing applications, it is used for analyzing the output power
through the fiber-optic sensor after transducer recognizes the interaction of
the analyte and the recognition unit (Gupta 2006).
1.3.4.3 Charge-Coupled Device

A CCD is a light-sensitive integrated circuit which is used to store and
display the data for an image. The working of CCDs involves the conver-
sion of each pixel (picture element) in the image to an electrical charge,
the intensity of which is related to the color in the color spectrum. These
are used as sensors in digital cameras and video cameras for the record-
ing and storage of steady and moving images. The CCD captures light and
transforms it to digital data, which is stored by the camera. The quality of
an image captured by a CCD depends on the resolution of the sensor. In
digital cameras, the resolution is measured in megapixels (or thousands
of pixels). Therefore, an 8MP digital camera can capture twice as much
information as a 4MP camera. The result is a larger photo with more detail.
These are used in the biosensors for the human-noninvasive monitoring
under clinical situations.
1.4 Sensor Performance Parameters

In this section, we shall discuss various static and dynamic parameters
which are used for the characterization of the sensor to evaluate its perfor-
mance. On the basis of these parameters, the sensor performance can be
enhanced which is greatly required for the advancement in the sensor tech-
nology. These are the following parameters:
• Sensitivity: Sensitivity is one of the most important parameter of the

sensor. It can be measured by calculating the change in the sensor
output signal with respect to unit change in analyte concentration
which is to be detected. It can also be evaluated from the slope of the
calibration curve, which is generally the plot between the measured
output signal and analyte concentration.
• Selectivity/specificity: Selectivity of the sensor corresponds to whether
the sensor responds either to a group of analytes or the sensor
responds specifically to a single analyte. The sensor is highly selec-
tive, if it responds only to the target analyte to be sensed in the pres-
ence of interfering analytes.
• Operating range: The analyte concentration range to which the sen-
sor operates is the most important parameter and it decides the sen-
sor’s applicability in the industry/hospitals, etc. Usually the sensors’
operating ranges are fixed because an increase in the sensor operat-

ing range decreases the sensitivity of the sensor.
• Linear range: The linearity range of the sensor defines the sensor’s oper-
ation with fixed sensitivity, which makes the sensor to be operated eas-
ily. This can be extracted from the calibration curve of the sensor by
fitting an ideal straight line. In other words, this tells us the analyte
concentration range up to which the output signal varies linearly.
• Limit of detection: The lowest analyte concentration which can be
detected by the sensor is defined as the limit of detection (LOD) of
the sensor. The evaluation of the value of LOD depends upon the
type of sensor fabrication and the detection system.
• Limit of quantification: Limit of quantification (LOQ) is also referred
to as the lowest analyte concentration which can be detected by the
sensor, considering the standard deviation in the output signal.
• Resolution: It is the lowest change in analyte concentration which can
be recognized by the sensing device. This is an important parameter
for the sensors in flowing streams.
• Response time: Response time of a sensor denotes the time taken by
the sensor to show a saturated output for a given change in the ana-
lyte concentration. Generally, it is defined as the time required by
the sensor to respond for the analyte detection.
• Repeatability and stability: As named, these terms correspond to the
sensor’s ability for reproducing its output response after characteriz-
ing the sensor up to certain cycles and maintaining its performance
for a specific time period. These parameters play the most important
role for the sensor to be commercialized.
• Hysteresis: It is the main characteristic of the sensor which decides
the difference in sensor’s response for an increasing and decreasing
analyte concentration. It is normally given as the percentage change
in the output response after the interaction and removal of the ana-
lyte from the recognition unit of the sensor. Zero percentage hyster-
esis refers to the exactly reversible nature of the sample.
• Shelf life: This parameter refers to the time period for which the
sensor will operate. If the sensor is not usable for a favorable time
period, then it affects the sensor’s preference.
1.5 Biosensor Classifications

An independent sensor incorporating a biological material in its active form
in the system, to interact with the analyte which is chemical/biological, is
generally considered as a biosensor. The design, selection, and application

of biosensors are critical since its interaction with the desired analyte to
be sensed is always affected by interferands. The application needs to be
performed in external environments, harsh atmospheres, laboratories and
health clinics, and even for in vivo and in vitro measurements in human body.
The analytes and the active biological elements may also need to be sterilized
in some cases, which consume time, high-cost equipments, man power, and
control measurements. However, the possibility of purifying these increases
the interaction of sensor with the analyte and provides better analysis.
The necessity of sensors in the area of health, environment, pharmaceu-
tical, agricultural, water monitoring and food quality, hygiene, drug and
medicine development, and dialectic cases is increasing. The sensor should
be easily available, economical, portable, easy to handle, commercialized
into a small module so that even laymen can use it for agricultural applica-
tions, food safety, and environmental monitoring. Thus, the processing and
the analysis should be performed in a smart way. In the case of biosensors,
especially, the analysis should be rugged to make the sensor extremely selec-
tive, sensitive, reliable, recoverable, and vigorous. This is where we can find
the difference between biosensors and bioanalytical schemes, which require
multiple processing and are of onetime use. For the use of biosensor as a tool
for diagnosis, in medical field such as cancer/tumor cell determination, the
sensor needs to be reproducible with capabilities of multianalyte detection
(Tothill 2009).
Biosensor works either in direct or indirect monitoring mode, depending
on the kind of reaction or analysis the sensor is intended to perform. In direct
monitoring mode, the analysis is done by monitoring the direct products and
reactions occurred due to the interaction of analyte (Gauglitz 2005), whereas
in the case of indirect monitoring mode, the analysis is performed indirectly
through the inhibitors or catalysts consumed for the reactions. The central
unit of every biosensor is the bio-recognition element synced with the trans-
ducer unit. The functioning of the biosensor depends on its recognition ele-
ment and transducer. Hence, biosensors are mainly classified on the basis of
their recognition element and transducer units. In the following sections, we
shall discuss the classification of biosensors and their importance for differ-
ent applications and measurements by correlating the recognition and trans-
ducer modes.
1.5.1 Classification Based on Transducer

The transducer part of the biosensor system is the unit, which provides
bidirectional signal conduct. The transducer part determines the signal
mode available at the output for analysis and readable by humans. There
is a predetermined relationship between the output and the input signal in
the biosensor system, and by calibrating the results from the analyzer one
can evaluate the measurements and changes. The transducer element in a
biosensor can be chosen according to the design and substrate of the sensor
probe. Thus, based on the transducer element, biosensors can be broadly
classified into five main types as electrochemical, optical, calorimetric,
mass-sensitive, and light-addressable potentiometric transducer (Patel 2002,
Reyes et al. 2003).
1.5.1.1 Electrochemical Transducer

Electrochemical types of transducers are the commonly used ones because
of their fast response, automation possibility, and compatibility with modern
technologies of fabrication, portability, and minimal requirement of power
(Thevenot et al. 2001). These sensors are usually used in point-of-care appli-
cations such as glucose sensor. In the case of electrochemical sensor, the
substrate is the electrode, which is chemically modified. This can be done
by modifying the electrode with ionic/conducting/semiconducting material
along with a chemical/biofilm (Durst et al. 1997). There are two electrodes,
one a working electrode which performs sensing and another a reference
electrode dipped in the electrolyte. Electrochemical sensors can be potentio-
metric, amperometric, conductimetric, and impedimetric.
An amperometric electrochemical sensor setup consists of working elec-
trodes (cathode and anode) and a reference electrode dipped in an electro-
lyte as shown in Figure 1.3. For a particular potential applied between the
electrodes, a chemical reaction occurs due to the oxidation or reduction
of electroactive species. The process of oxidation or reduction generates
current in correlation with the concentration gradient of the electroactive
species/analyte which is measured by an ammeter. For the sensing pur-
pose, the receptor is immobilized over one of the electrodes. When analyte
is added into electrolyte, the current starts flowing with the electroactive
species toward the electrodes. In the presence of receptor, the analyte binds
with the receptor and the conductivity of the receptor unit changes. Since
the receptor is immobilized over the electrode, the conductive properties of
the transducer surface change. This results in the change in current flow-
ing through the electrode setup, which is analyzed by the ammeter. Thus,
by observing the change in current, before and after adding the analyte,
in ammeter, one can detect the presence of analyte. Electrodes are gen-
erally made up of metals/steel/carbon that are conductive in nature and
are modified according to the application. The bio-recognition element
and the immobilization methods with matrix can be varied accordingly,
which may also use an electron mediator or a selective/discriminative
membrane. Amperometric sensors are fast, more sensitive, and exact than
other types in electrochemical system. The calibration curve of these sen-
sors is, in general, linear. The first biosensor using amperometric trans-
ducer was designed for glucose monitoring based on the consumption of
oxygen (Updike and Hicks 1967). This kind of amperometric sensor, which
performs analysis based on the consumption of indicator or formation of
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own I trembled as I said it—I told him I was coming to see you here,
that I would no more forsake you in your trouble than your mother
would have done. She would have clung to you in your darkest hour
because she loved you, and that I did not love you less.”
“And he did not forbid you?” cried the young man in amazement.
He knew Rupert Morrice so well, of a nature singularly kind and
generous, but hard as flint to evil-doers, to those who betrayed his
trust.
“No, he did not forbid me, Dick, and I am almost as amazed as
you are, but I think that reference to your mother softened him. It
was a long time before he spoke, and then his words came very
slowly. ‘If I allow you to do this, at any rate for the present, till I have
thought matters out further, will you give me your solemn promise
that you will only play the rôle of consoler, that you will do nothing
rash?’ Of course I knew what he meant, that we might get secretly
married. So I gave him that promise, Dick; do you blame me?”
“A thousand times No, my darling,” cried Croxton, as he took her
in his arms and kissed away the tears on the sweet face. “And
considering what he believes me to be, nay more, what he is sure I
am, I cannot but marvel at his giving his consent.”
For a long time the young people talked together, and all through
their conversation the one thought uppermost in Rosabelle’s mind
was that her lover should take steps to clear himself, that he should
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“If you do not act, I shall act myself,” she told him finally.
The young man listened attentively, and hope and resolution
began to stir in him. He had been so stunned by the damning nature
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benevolent protector, that he had been crushed almost into
insensibility, into a benumbing of his faculties. But, as the girl spoke
in her bright, incisive way, the clouds about his brain seemed to melt.
He seemed to see himself rehabilitated, able to prove to those whom
it concerned that he was the honest man they had always believed
him to be.
“The question is how to go to work,” he said gravely. “Mr. Morrice
is right when he says that to call in Scotland Yard might lead to
disastrous consequences. But we could employ a private detective
to probe the mystery to the bottom. Even if he could not lay his
hands on the actual thief, he might be able to prove my innocence.”
Rosabelle caught eagerly at the idea. “And where can we find the
sort of man we want?”
“One of the cleverest is Gideon Lane; his office is in Shaftesbury
Avenue. I know him a little, and Mr. Morrice knows him too. We
employed him to watch a suspected clerk in our office, and he
trapped him very cleverly.”
“Would it cost much to employ him?” asked the girl anxiously. She
knew that Richard’s capital, like her own, was very small, and it was
hardly likely that Morrice would spend any money on a case he had
already pre-judged. It was not possible for her to help, for her uncle
was her trustee and not likely to allow her to adventure a penny in
such a cause.
But Croxton’s small amount of capital was entirely under his own
control, and now that he was recovering from his despairing mood,
he was fired with the desire to establish his innocence, and had no
hesitation in employing some of it for the purpose.
After a great deal of discussion as to the initial steps to be taken, it
was decided that Rosabelle should visit the detective, tell him the
whole facts, and commission him to undertake the investigation on
her own behalf. Richard would give her a brief letter of introduction to
Gideon Lane, and furnish her with money to pay a preliminary fee.
The enthusiastic girl did not allow the grass to grow under her feet.
Two days later she was seated in the waiting-room of the small suite
of offices in Shaftesbury Avenue. She had sent in her letter of
introduction and was waiting to be summoned to the presence of the
well-known detective who was, fortunately for her impatience,
disengaged. He was not many seconds reading the letter, but it
seemed hours before the restless Rosabelle saw the inner door
open, and was asked by a smart young typist to step in.
Mr. Gideon Lane rose to receive her, a tall, good-looking man with
nothing particularly remarkable about his appearance; with his clean-
shaven face and strong, resolute expression he might have been
taken for an actor, there was certainly nothing about him to suggest
an unraveller of mysteries. The most striking features in an
agreeable countenance were his eyes, which were piercing and
brilliant.
“I remember Mr. Croxton perfectly,” said the detective. “He was the
confidential secretary of Mr. Morrice, and struck me as much above
the ordinary young man in intelligence and quickness of perception. I
hope he is quite well,” he finished politely.
This remark gave Rosabelle an easy opening. “He is quite well in
health, Mr. Lane, but exceedingly unhappy, lying as he is at the
moment under the stigma of a terrible accusation.”
Mr. Lane gathered from these serious words that the girl had come
upon a grave errand. His face reflected her concern at once.
“I am very sorry to hear it, Miss Sheldon. I took rather a liking to
the young man, he seemed so open and frank. Well, please tell me
all the details, I take it you want my assistance in the matter. And
please conceal nothing from me, if you want me to give you of my
best. Let me know everything that tells against him, you will naturally
inform me of everything in his favour.”
The shrewd man of the world divined immediately that there was a
close bond between this charming girl and the accused man, and he
put her at once at her ease by adding: “I need hardly tell you that
what you say will never be divulged; you are as safe with me as if
you were in the confessional.”
He had a very ingratiating manner with him, this calm, self-
possessed man who looked more like an actor than a detective.
Rosabelle felt very much at home with him, and at once launched
forth in her narrative of the details of that eventful morning, as they
had been told her by her lover.
Mr. Lane listened to her attentively without interruption. He judged
it best to let her tell her story her own way, more particularly as she
told it very well, without redundance or repetition. His questions
would come later.
When she had finished, he sat silent for some time, while the girl
regarded him anxiously. “It is, of course, too early for you to form any
opinion?” she asked in a faltering voice, feeling the prolonged
silence somewhat of a strain upon her nerves.
He shook his head. “A great deal too early, Miss Sheldon. Of
course, it is easy to say at first blush, upon the evidence before us,
those articles could only have been abstracted by one of two
persons, Mr. Morrice or his secretary.”
“And it would be absurd to think that my uncle stole his own
property,” cried the girl swiftly.
A rather non-committal smile illumined the calm face of the
detective. “From your point of view, it would be absurd, as you most
rightly say. From mine, it would be so very difficult to discover a
plausible motive for such an act.”
She could not follow him in this subtle explanation, and waited in
silence till he began to put certain questions to her. First, with regard
to the servants, would she give him full particulars of their number,
the nature of their duties, their length of service and so on?
She supplied him with the requested information. He entered all
this in a private notebook, in a shorthand of his own invention which
nobody could read but himself.
What did the family consist of? was his next question.
“My uncle and aunt, Richard Croxton and myself. Two other
people came to the house who were practically of the family, Sir
George Clayton-Brookes, my aunt’s brother-in-law, and young
Archibald Brookes, his nephew and the son of my aunt’s sister.”
These particulars went into the notebook. “I have heard of Sir
George, he is well known on the turf, and reputed to be a man of
substance. I know nothing of the young man. Has he means of his
own, or is he dependent upon his relatives?”
“Dependent upon Sir George, I believe,” answered Rosabelle. “We
have always understood his uncle makes him a handsome
allowance, and will leave him his property.”
Mr. Lane asked a few more questions and then closed his
notebook. “Well, Miss Sheldon, that is as far as we can go at
present. Before I start, I must visit the scene of operations and take a
look at this wonderful safe. I take it that will not be easy to
accomplish without Mr. Morrice’s knowledge and permission. Is he
likely to refuse it?”
Rosabelle, needless to say, was a little dismayed. He had refused
to call in Scotland Yard, would he peremptorily refuse admission to a
private inquirer?
She hazarded her fears to Mr. Lane, who thought that he would
yield in the matter. The fact that Richard Croxton was prepared to
break into his small capital for the purpose of establishing his
innocence, should make a favourable impression upon Mr. Morrice,
however firmly he believed in the young man’s guilt. If Morrice
obstinately refused, he would be forced to revise his opinion of that
gentleman, although he was too diplomatic to say as much to
Rosabelle.
“I will tell you the principal object of my visit, Miss Sheldon. The
theft would have to be committed in a great hurry, and there are sure
to be finger-marks on the safe. I want to take a photograph of them.
If Mr. Morrice does refuse, for reasons sufficient to himself, I shall
have to get a photograph of them somehow, and in this I dare say I
shall have to avail myself of your co-operation.”
He smiled a little as he spoke. It was not the first time by many
dozens that he had gone in at the back door where he had been
refused entrance at the front, or obtained information he required in
spite of every obstacle being put in his way.
Rosabelle was quite sure she understood what he was driving at.
She would have dared anything for her lover, and if it was a question
of smuggling Mr. Gideon Lane into her uncle’s room while he was in
the city, her woman’s wit, sharpened by her love, would find a way.
“Now we will not waste time,” said the genial Mr. Lane as the
excited girl rose to take her leave. “Pending the obtaining of your
uncle’s permission to do the thing openly, I want you to co-operate
with me in a little matter. Pay Mr. Croxton a visit as soon as possible
and get him to give you an impression of his fingers. If you tell him
what you want it for, he cannot refuse.”
“But, of course, he will not refuse,” cried the girl a little indignantly.
“Would he have let me come to you if he was not prepared to face
the ordeal? And if you find, as you will, that the finger-marks on the
safe are not his, that will establish his innocence once and for all, will
it not?”
Mr. Lane seemed a trifle embarrassed by the question. “It will go a
long way,” he said, speaking with some hesitation.
“Why not the whole way?” demanded Rosabelle, and her eyes
flashed a little.
“Miss Sheldon, it is better you should not ask me too many
questions till we are more sure of our ground. We experts require a
great deal of evidence before we venture to say of any accused man
that he is absolutely innocent or absolutely guilty.”
“But if the finger-marks are proved not to be his, how can he be
guilty?” she cried obstinately.
“You force me to say what I would rather leave unsaid. But our
investigations would not be very useful if we refused to weigh not
only every probability, but also every possibility. You say that your
uncle firmly believes in this young man’s guilt, although he loved him
and treated him like a son. If he still maintains that belief, is it not
open to him to say that if Richard Croxton was not the actual thief,
he was an accomplice or an accessory? How otherwise could the
actual thief have got the necessary knowledge of that safe’s
complicated mechanism? Please understand I am not advancing this
as my own opinion, but as one that might be entertained.”
And for the first time poor Rosabelle began to see how very hard
was the task before them. The tears came into her eyes. “Oh, Mr.
Lane, what will be wanted to prove his absolute innocence? I see too
clearly the terrible difficulties in our way.”
The great detective spoke very gravely. “The surest way of proving
Mr. Croxton’s innocence is by discovering beyond any possibility of
doubt the person who opened that safe, and proving that that
person, whoever it may be, had no connection with him. To that point
my investigations will tend, with what results it is impossible for me to
foresee.”
Mr. Morrice gave his permission for the detective’s visit more
readily than Rosabelle had hoped. His attitude towards young
Croxton now seemed to be more one of sorrow and disappointment
than of the deep anger he had at first displayed. But he expressed to
her his sense of the futility of the task on which she was engaged.
She thought she knew what was passing in his keen and analytical
mind. Croxton was playing a game of bluff, perhaps for the purpose
of establishing himself firmly in the esteem of his sweetheart. And if
the finger-marks were those of somebody else, he would fall back on
the theory that Gideon Lane had already anticipated.
With Richard, her task was easy. He gave an impression of his
fingers without a moment’s hesitation, and Rosabelle carried it to
Lane with a certain sense of triumph, which would have been
complete but for those last damping words of the cautious detective.
In due course the visit was paid to the house in Deanery Street;
Rosabelle and her uncle were present. Sure enough in addition to
the recent finger-prints of Morrice and young Croxton, there was a
third set, equally recent.
The development of the photographs proved that Croxton’s finger-
prints were totally different from the third set. Lane announced his
intention of taking them to Scotland Yard in order that a search might
be made amongst their voluminous files.
His investigations on this subject completed, Lane dispatched a
brief telegram to Rosabelle asking her to call at his office. A few
minutes after its receipt, she was seated in his room feverishly
awaiting his news.
“It promises to be a deeper mystery than I thought, Miss Sheldon.
There has been some very clever and deeply thought-out work here.
I have identified the finger-prints, they are those of a well-known
professional thief named Thomas, known amongst his confederates
as ‘Tubby’ Thomas. He is an expert safe-breaker, the cleverest in
England.”
The girl’s eyes sparkled. “An expert safe-breaker!” she repeated
joyfully. “Does one want to pursue the inquiry any further? Is it not
obvious who was the thief?”
But the next moment came the slow words which fell like ice on
her heart.
“Unfortunately, the mystery is deepened, not solved. The finger-
prints are those of ‘Tubby’ Thomas, for finger-prints never lie. But
‘Tubby’ Thomas himself has for the last two years been serving a
sentence for a similar offence in Dartmoor, and he is still there.”
CHAPTER V
ROSABELLE AND LANE CONFER
D AZED as she was, cast in a moment from a feeling of elation

into one of bitter disappointment, she saw the point at once. If
the criminal known as “Tubby” Thomas was safe under lock and key,
he could not have been the thief. They were as far from the solution
of the mystery as ever, in spite of those tell-tale finger-prints which,
according to orthodox belief, never lied.
Gideon Lane was bitterly disappointed too, but he had suffered so
many checks in his time that he never allowed his fortitude to desert
him. When he discovered those finger-prints he really thought the
game was in his hands, and that, with the aid of Scotland Yard, he
could put his hand on the actual thief, as he could have done had
they been those of a criminal actively pursuing his nefarious career.
But the incarceration of the man Thomas provided an impasse.
Narrowing the issue to the only two men who were supposed to be
acquainted with the complicated mechanism of this wonderful safe,
he had thought very deeply, twisting and turning about in his keen
and alert mind the possibilities that suggested themselves.
Taking the young man himself first. According to the flattering
report of Rosabelle, he led a perfectly blameless and open life. In his
habits he was temperate, almost abstemious, he never touched a
card, he never betted, the only gambling habit he indulged in was to
take a ticket in a couple of club sweepstakes. But, of course,
Rosabelle’s report was sure to be coloured a little on the favourable
side. There are plenty of young men who lead double lives; models
of discretion and decorum to all appearances, but secretly addicted
to ruinous and discreditable vices which are only brought suddenly to
light by some accident or fatal false step.
This young man might be one of these. He might be hard pressed
for money, the victim even of some blackmailer who had become
possessed of a terrible secret in his double life, and had risked all his
bright prospects on the chance that Morrice would disbelieve the
evidence of his senses, and accept his bare denial that he was
innocent, in spite of the damning evidence against him.
But if he was clever enough to scheme out such an artfully-
planned robbery, either alone or with the aid of a confederate, would
he not be clever enough to see that scrupulous honesty and fidelity
to his employer was the best policy? For Morrice, according to
Rosabelle’s account, had treated him like a son; there was little
doubt that he intended to take him into partnership at an early date,
and would leave him a considerable slice of his vast fortune. There
was no doubt of his wealth, for, by common consent, he was reputed
to be amongst the half-dozen richest men in England.
Then there was no doubt that the two young people were lovers.
Would a man, capable of a moment’s sane thought, put in certain
jeopardy his chances of happiness with this charming and lovable
girl?
But then, of course, crimes would never be perpetrated if the
criminals could foresee all the consequences likely to flow from their
yielding to sudden impulses. At the fatal moment they appeared to
be driven forward by some blind force which, for the moment, they
were unable to fight against. And so it might have happened in the
case of this young man, who, according to Rosabelle’s testimony,
had led such a regular and blameless life.
Turning his attention to the other of the two men, Rupert Morrice
himself, the detective found the situation one of greater complexity.
Strange as it may sound, men have robbed themselves before now
and done their best to fix the guilt upon others, from more than one
sinister motive. For instance, a man knowing himself to be on the
verge of bankruptcy might, in desperation, purloin some of his own
property to put it in a safe place beyond the reach of his creditors. In
the case of this wealthy financier, whose credit stood so high, such a
theory might be at once dismissed.
At first blush, the refusal to apply to Scotland Yard might seem a
trifle suspicious, might suggest that he had a personal interest in
stifling independent investigation. But when one considered the
unusual circumstances, the action seemed only a natural one.
According to Rosabelle’s statement, Morrice had treated the
young man as a son; not only had he a great affection for him, but
that affection had been accentuated by the elder man’s passionate
love for the mother. However deep his belief in his guilt, a father
does not hand over a son to be dealt with by the stern processes of
justice. He may dismiss him from his house, he may refuse to hold
further intercourse with him, but he shields him, where possible, from
the fatal consequences of his rash act.
There was, however, one point on which he wished to be assured,
and which caused him to put a certain question to the girl.
“I am going to ask you something, Miss Sheldon, not, believe me,
from any spirit of impertinent curiosity, but because it is essential that
I should be acquainted with every little fact. I am assuming that your
interest in Mr. Croxton arises from a warmer feeling than that of mere
friendship. Am I not right in saying that there is a close bond
between you; that, to put it in plain words, you are lovers?”
Rosabelle admitted quite frankly that Lane was right in his
surmise.
“Now for my next question. Did Mr. Morrice know of this
understanding between you, and if so, did he approve of it?”
To this the girl’s answer was equally frank. Up to the day of the
robbery she could not have been absolutely certain that her uncle
did know of it, although she was pretty sure he did. Their interest in
each other was so openly displayed, that it was almost impossible it
could have escaped his observation. If he had disapproved, he
certainly would not have hesitated to express his disapproval, being
a man of the most straightforward character, who never scrupled to
express what was in his mind, or take drastic action when he judged
it necessary.
“All doubt, however, on this point was removed by what he said to
Richard on that terrible morning,” Rosabelle went on in a voice that
trembled a little. “After overwhelming him with his anger at what he
believed to be his baseness, he told him he knew we were attached
to each other, and that he would have put no obstacles in our way. It
was really as I thought. Richard was always a little dubious as to
what his attitude might be, while I never had the slightest fear. We
were both so very dear to him that I was always sure our marriage
would have given him the greatest pleasure.”
The detective considered her reply carefully, as was his invariable
custom. He never accepted any statement without probing it very
deeply, none knew better than he the futility of jumping to rash and
hasty conclusions.
“There would seem to be some reasonable ground for Mr.
Croxton’s doubts in the matter,” he said very quietly. “Kind and
generous as Mr. Morrice was to him, there was no actual blood-tie
between them; you tell me the young man had practically no money
of his own, that his future depended entirely on a continuance of his
benefactor’s favour. You, on the other hand, are a near relative and it
is to be assumed that your uncle will leave you a considerable sum.
It would be a very natural thing that he should have different views
for you, should have wished you to look a little higher than one who,
after all, was not your equal in anything but birth. At any rate, it is
what the ordinary person might think, of course; Mr. Morrice may be
an exceptional man of liberal independent views.”
“Oh, but that is just what he is,” cried the girl warmly. In spite of her
fervent belief in her lover, and perhaps a little natural resentment
against her uncle for his obstinate presumption of Richard’s guilt,
she loved him very dearly and thoroughly appreciated his sterling
qualities.
“That is just what he is, Mr. Lane,” she repeated. “Rich as he is,
hard as he works to make himself so, he does not love money for its
own sake or value the possession of it in others. One or two of his
closest friends are poor men, and he is happier in their society than
in that of millionaires like himself. He loves his business and his
work, it is true, but more for the mental excitement and stimulus they
bring than for their pecuniary results. And he doesn’t attach much
importance to birth or what the world calls position. At heart, I believe
he is a good bit of a democrat.”
“If a millionaire can be truly a democrat!” suggested Mr. Lane with
a smile. “Anyway, if he is one, there must be a good many
reservations.”
The girl’s replies to his questions had rather disposed of a
somewhat fantastic theory that had formed itself in rather nebulous
shape in his astute brain, accustomed to weigh all sorts of
possibilities and probabilities, to search for unusual and far-reaching
motives. Had Morrice engineered this theft, not for the ordinary
sordid reasons, but with the object of fixing upon the innocent
secretary a stigma that would effectually remove him from his niece’s
society? But then again, a man who could in cold blood conceive
such a scheme would be more than the vilest criminal. It would be
impossible that one of such good repute, for even his enemies and
rivals credited Morrice with the highest integrity, should stoop to such
sinister methods.
“Well, Miss Sheldon,” he said as the interview drew to a close, “I
will not disguise that I am very disappointed with the result of my visit
to Scotland Yard. When I found those strange finger-marks on the
safe, I thought we were on the right track. Now, I have got to start
again from the beginning, and I am afraid it will be a long time before
I shall make any considerable headway. I shall do my best, but it
may be that in the end I shall be beaten. I think you said you would
be going abroad very shortly.”
“Yes, we start for Mürren a week before Christmas for the winter
sports. I was so looking forward to it, but now——” The girl’s voice
faltered and she could not finish her sentence.
“I quite understand,” said Mr. Lane soothingly. “All the same it will
be better for you to get away for a time from these painful
associations. I will, of course, keep in touch with you to the day of
your departure, and communicate to you anything of importance. If
you don’t hear from me, you will know that so far I have nothing to
tell you. You will, of course, acquaint Mr. Morrice with the rather
puzzling information about the man Thomas, that while the finger-
prints are undoubtedly his, he is and has been for the last two years
in prison.”
It all seemed very hopeless, she thought, as she rose to leave. It
was useless to ask Lane if he had formed any theory; she had seen
enough of the man to know that he would not say a word till he felt
himself justified in speaking.
“One little thing before you go, Miss Sheldon. Will you kindly let
me know your aunt’s maiden name, and, if you possess them, any
particulars of her family.”
Rosabelle did not know much beyond the fact that she was a Miss
Larchester; that her sister, no longer living, the mother of Archie
Brookes, had married a younger brother of Sir George. She was not
quite sure but she fancied that, as a girl, Mrs. Morrice’s home had
been in Sussex, but she did not know in what part. The lady very
seldom alluded to her past life. Her Christian name was Lettice.
Mr. Lane entered the scanty information in his notebook, then,
after Rosabelle’s departure, he rang up White’s Club and inquired for
a Mr. Sellars. In a few moments this gentleman was speaking to him.
“Good-day, Mr. Sellars. I should be obliged if you would come
round to me as soon as convenient.”
The reply was that the owner of the name would at once put
himself in a taxi and be there in a few minutes.
CHAPTER VI
LANE ENGAGES AN ASSISTANT
M R. SELLARS, Reggie Sellars as he was known to his intimates,

was a tall, good-looking young man of about thirty, of the
aristocratic type, with aquiline features and an elegant figure.
Following no settled occupation or profession, he formed one of that
numerous brigade of men-about-town who belong to good clubs,
frequent respectable society and always seem to have plenty of
money for their personal wants, although nobody knows the exact
source of their incomes, or how they contrive to present such a good
appearance.
These men are usually very scrupulous in money matters, pay
their bets promptly when they lose and expect to be paid as promptly
when they win, are never behindhand in a club subscription, liberal
but not ostentatious in their tips to waiters. Many of them, in fact
most, have a small annuity which forms the nucleus of their income;
how the rest of that income is earned is often a puzzle to even their
most intimate friends.
Mr. Sellars was one of a large family, some twelve in all, sons and
daughters. His father had left capital bringing in about fifteen
hundred a year to be divided amongst this numerous offspring. This
brought Reggie in the modest competence of about a hundred odd
pounds a year. He dressed very well, and his tailor must have taken
more than that. It was obvious, therefore, that he had a knack of
picking up money somehow and somewhere, as he belonged to
several clubs, frequented fashionable society, and was by no means
an anchorite in his tastes.
As a matter of fact, he lived on his wits, using the expression in a
perfectly respectable sense. He furnished gossip to a well-known
Society newspaper for which he received a liberal remuneration; he
was a scientific backer of horses, he played a first-rate hand at
bridge, sometimes he got a handsome fee for initiating some
nouveau riche into the mysteries of fashionable life. Since his
acquaintance with Mr. Gideon Lane, he had often been useful to that
gentleman, and had been paid well for his services.
They had met at a Bohemian club to which both men belonged, for
Reggie Sellars, although of very good family and an aristocrat by
instinct and connection, was by no means exclusive, and was
equally at home in Bohemia and Mayfair.
At first Lane had not been attracted to the young man, whom he
regarded as the usual type of lounger who led a life of aimless
pleasure, a mere idler with whom he was not likely to have anything
in common. And, truth to tell, although in a certain way he was one of
the shrewdest fellows alive, Sellars’ good-looking countenance did
not furnish any striking evidence of mentality or strenuous impulse.
But one night in the smoking-room the two got into a conversation
on the subject of criminals and criminology, and Lane found that this
seemingly idle, pleasure-loving young man, with apparently no
thoughts beyond the race-course and the bridge-table, displayed a
keen knowledge and a swift power of deduction that astonished him.
Lane had a considerable clientèle amongst persons high up in the
social scale, he frequently wanted to obtain special information about
people belonging to or moving in fashionable circles. Into such
quarters he was unable to penetrate himself for obvious reasons.
Here was a man just fitted for the job, keen, quiet, quick in resource;
a man, in short, disguising a considerable mentality under a most
deceptive exterior. Lane suggested that there was certain work in
which his previous knowledge and facilities of approach could be of
material assistance to him. Mr. Reginald Sellars, the good-looking
young man-about-town, jumped at the proposal, and Lane had to
confess that, in his own line, he had never possessed a more
competent lieutenant.
He was just the man for the Morrice job, or at any rate one
particular portion of it, and that was why the busy and brainy
detective had rung him up to-day.
“Not been very long, eh, Lane?” was the young man’s greeting as
he entered the private room. “Always ready for business, you know,
for anything that brings grist to the mill. I hope you’ve got something
good for me.”
At his fashionable clubs, in the society of his aristocratic friends,
he cultivated a rather languid manner. When he talked to practical
people like Lane his tone was brisk, his whole manner alert.
The detective went to the point at once. “Of course, you know of
Rupert Morrice, the big financier, most probably you are personally
acquainted with him?”
“Known him for years, he was rather a pal of my father’s, used to
give him a good tip now and then for his investments,” was the
answer. “Can’t say I’m one of the intimates of the house, but always
get a card for their big things, have been asked twice, I think, to fill
up a dinner-party. What’s up?”
But without answering his question, Lane asked one himself. “We
all know the man’s story, that is public property. But what about Mrs.
Morrice; do you know anything about her antecedents, her family,
her history, before she met her husband?”
Sellars shook his head. “I’ve never heard, I don’t think anybody
has. A very charming woman, well-bred and all that, does the
honours perfectly, but never seems to talk about herself as most of
her sex do. The only thing I can remember is that some few years
ago a nephew was introduced, a young chap named Archie Brookes,
who was also a nephew of Sir George Clayton-Brookes who is as
well-known in London as the Monument. Her sister married his
younger brother, we were told.”
“You don’t know her maiden name?”
“No, but that of course can easily be got at Somerset House,” said
the bright young man who had proved such an able colleague.
“Of course, I know that, but we need not go there. I have got the
name, a rather uncommon one. She was a Miss Lettice Larchester,
and I believe she hails from somewhere in Sussex.”
“And you want me to find out all about her before she became
Mrs. Morrice, eh? He met her and married her abroad, I suppose you
know that. He was awfully gone on Mrs. Croxton, the mother of that
young chap whom he practically adopted and who acts as his
secretary. It is said he remained a bachelor for years because of
her.”
Reggie Sellars’ knowledge of the annals of the people who moved
in certain circles was of the most exhaustive nature. And he had a
memory like a vice; he never forgot a fact or a date, and never
confused one history with another. He was certainly a most
deceptive person. To look at him you would never imagine he would
take the slightest trouble to acquire any knowledge that was not
strictly necessary for his own immediate purposes.
“Yes, I want you to find out all you can about her; of course you will
make your inquiries very discreetly. But, there, I need not warn you
of that. You are always discreet.”
And in truth he was. He could pursue the most delicate
investigations without giving himself away for a second.
“Well, now, you haven’t given me an inkling of what’s up yet, and
you know I’m not fond of working in the dark. Why this sudden
interest in Mrs. Morrice’s past?”
Lane was not addicted to telling more than he could help, for
secrecy had become an ingrained habit with him. But the young man
was a bit touchy on some things. He was especially so on the point
that perfect confidence should be reposed in him, and it must be
admitted that that confidence was never abused. He was a perfectly
honourable young fellow, and his word was better than the bond of a
good many people.
So Lane told him the salient details of the robbery in Deanery
Street, ending with the remarkable discovery of the finger-prints of
“Tubby” Thomas, and the incarceration of that accomplished
criminal.
The quick mind of Sellars speedily grasped the complicated nature
of this puzzling case. “By Jove, it wants a bit of thinking out, doesn’t
it, Lane? In the meantime, according to your invariable custom, you
are suspecting everybody, including Mrs. Morrice; the secretary, of
course, and Morrice himself, and naturally the Brookes’s, uncle and
nephew.”
Lane smiled. “I intend to know everything I can about every one of
them. I exclude the servants, it is too deep a job for any of them.”
“And what about that pretty girl, the niece, what’s her name—eh,
Miss Sheldon? You’ve got your eye on her, of course?” He spoke in
rather a joking manner, for he often rallied Lane on his tendency to
reverse the usual principle of British law and believe everybody to be
guilty till his innocence was fully established.”
“She is a very charming young lady,” replied the detective a little
grimly, for he did not relish being chaffed. “But I shall certainly not
exclude her from the scope of my investigations if all others fail. Well
now, look here, Mr. Sellars, I expect it will take you a little time to get
at Mrs. Morrice’s history. What do you know about this Clayton-
Brookes and his nephew? The uncle is a great racing man, I
understand, and you are amongst the racing set.”
“I know Sir George just a little, we nod to each other when we
meet, but I don’t think I have exchanged half a hundred words with
him in my life. Archie Brookes I know about as well. But I can tell you
this, he is not popular; most people think him a bit of a bounder. Do
you want me to investigate in that quarter too?”
“Yes, I wish you to find out all you can. I want you to discover
particularly what is known about the young man’s father who,
according to what we are told, married Mrs. Morrice’s sister.”
“Right, it shall be done,” replied Sellars. “Now, as I have said, I
don’t know either of the men well, and I can’t get any information
from them. But I do know pretty intimately a man who is a great pal
of Sir George; he’s a member of White’s, a good, garrulous sort of
person, and he’ll talk by the hour when you once get him started. I’ll
tap him as soon as I can get the chance. He’s much older than I, of
course, but we are rather pals, and I’ll make him give me what I
want.”
Lane did not possess a very keen sense of humour, his calling did
not greatly encourage it, but he was a bit tickled by the gusto with
which this remarkable young man, who hid his talents so
successfully under that indifferent exterior, set about the task of
extracting information from his numerous friends and acquaintances.
For it was one of his greatest assets, moving as he did in so many
various circles, that if he could not get what he wanted directly, he
could always do so indirectly. Here, for example, although he did not
know Sir George very well, he was more than intimate with that
gentleman’s great friend, whom, of course, he could pump with
greater freedom than Sir George himself. Presently he took his
leave, promising to let Lane know the result of his investigations at
the earliest moment.
He appeared a couple of days later. “He rose to the fly beautifully,”
he said in that brisk voice which he always assumed when he was
engaged on strict business. “He has got it all pat. Sir George had a
younger brother Archibald, a bit of a rolling-stone. He couldn’t make
good here, so his family packed him off to Australia to try what a
change of climate might do. He didn’t do very well there, but he
didn’t come back. He married—but my friend doesn’t know the
maiden name of his wife; Sir George had either never mentioned it,
or he had forgotten it. Anyway, there was one child, the boy Archie,
named after his father. The mother died a few years after his birth.
The father died later in Melbourne. When the young one was grown
up, Sir George sent for him to come home, and adopted him. There’s
the whole history cut and dried for you.”
“And very lucidly told too,” said Mr. Lane approvingly. Sellars knew
him well now, and he inferred from the careful way in which he
entered the details in his notebook that he attached great importance
to the information. So he did, much more than the young man
guessed; this he was to learn later on.
To be a really great detective a man must have a certain amount
of inspiration and imagination, and Lane possessed both these in a
remarkable degree. While ruminating over the various problems of
this puzzling case, one of these flashes of inspiration had come to

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Optical Sensors for Biomedical

Diagnostics and Environmental

Acoustic Sensors for Biomedical Applications Nilanjan

Nanosensor Technologies for Environmental Monitoring

Smart Sensors for Environmental and Medical

Simulations and Optical Diagnostics for Internal

An introduction to distributed optical fibre sensors

Sensors, Algorithms and Applications for Structural

Plastic Optical Fiber Sensors: Science, Technology and

Novel Sensors and Sensing First Edition Jackson

Banshi Dhar Gupta

© 2018 by Taylor & Francis Group, LLC

No claim to original U.S. Government works

Printed on acid-free paper

International Standard Book Number-13: 978-1-4987-8906-6 (Hardback)

Library of Congress Cataloging-in-Publication Data

Visit the Taylor & Francis Web site at

2.3.1.1 Long-Range SPR...................................................... 55

4 Nanostructured Sensors............................................................................. 103

4.2.1.1 Chemical Methods................................................ 109

5 Semiconductor Metal Oxide Sensors...................................................... 165

5.5 Biosensing with SMOs...................................................................... 189

6 Molecular-Imprinting-Based Sensors..................................................... 197

7 Summary and Future Outlook.................................................................. 227

This book is an effort to bring a comprehensive book on optical sensors for

healthcare in this chapter. Chapter 2 introduces the basic concepts of reso-

Banshi Dhar Gupta

Banshi Dhar Gupta

Sruthi Prasood Usha earned her BTech in electronics and communication

in 2014. She has published 12 research papers in international journals

Measurements of various physical, chemical, and biological parameters/

1.1 What Is a Sensor?

1.2 Need of Sensors

the applications of sensors for biomedical, clinical, environment, and food

advancement exploiting the enzymes, antibodies, and microbes in combina-

1.2.2 Gas Sensors

1.2.3 Chemical Sensors

1.3 Sensor Components

diagram showing the sensor components with possibilities in their selection

1.3.2 Recognition Unit

1.3.4.2 Optical Power Meter

1.3.4.3 Charge-Coupled Device

1.4 Sensor Performance Parameters

• Sensitivity: Sensitivity is one of the most important parameter of the

operating ranges are fixed because an increase in the sensor operat-

1.5 Biosensor Classifications

generally considered as a biosensor. The design, selection, and application

1.5.1 Classification Based on Transducer

1.5.1.1 Electrochemical Transducer

D AZED as she was, cast in a moment from a feeling of elation

M R. SELLARS, Reggie Sellars as he was known to his intimates,

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