Boyono Tiny ELISA Absorbance 96 Reader ENZ - Manual
Boyono Tiny ELISA Absorbance 96 Reader ENZ - Manual
Boyono Tiny ELISA Absorbance 96 Reader ENZ - Manual
Rev 102320
Instructions for Use
Absorbance 96
Software Version 1.1.0
I Ensure that the manual is accessible at all times and is read and un-
derstood by all persons operating the instrument. Keep this manual in
a safe place so that you can access it when necessary.
I The latest version of this manual (instructions for use) is available via
the Absorbance 96 App. Upon request, a paper version can be
shipped within 7 business days. Please use the contact form on the
website.
This user manual may only be used according to its intended purpose. It may not
be reproduced, changed, or translated in another language without prior written
consent of Byonoy GmbH.
2
TABLE OF CONTENTS
4 Operation ...........................................................................................................................................11
6.1 Maintenance................................................................................................................ 23
6.2 Cleaning ........................................................................................................................ 24
6.3 Technical support ...................................................................................................... 26
6.4 Repairs .......................................................................................................................... 26
8 Guarantee ....................................................................................................................................... 28
3
General information
1 General information
Any commissioning or handling of the instrument requires precise knowledge and notice of
this manual. The device is intended for the described use only.
Warning:
!! This is a warning that indicates an immediate or potentially dangerous situation,
which, if not avoided, will result in death or serious injury.
Caution:
! Caution indicates a potentially hazardous situation, which, if not avoided, could
result in minor or moderate injury.
Information:
I This is a piece of information indicating certain characteristics that must be ob-
served.
Liability for the function of the device shall in any case pass to the owner or operator if the
device is improperly maintained, repaired, or modified by persons who do not belong to the
authorized service personnel, or if it is handled in a way that does not comply with its in-
tended use. The service and operation of the product must be in accordance with this man-
ual. Byonoy GmbH shall not be liable for damages resulting from non-observance of the in-
formation above. Warranty and liability conditions of the terms of sale and delivery of Byonoy
GmbH are not extended by the information above.
Warning
!! Incorrect cleaning of the instrument can reduce its service life and can cause
damage to the instrument. Please note the explanations and warnings in chapter
6.2.
!! To avoid the risk of electric shock, do not expose the instrument to rain or ex-
cessive moisture and do not touch the instrument or USB cable with wet hands.
4
General information
Information
I Follow all safety instructions on the device and in the attached documents.
The symbol on the product or its packaging indicates that this product is not to be treated
as normal household waste. It must be disposed of at a collection point for the recycling of
electrical and electronic equipment. By contributing to the correct disposal of this product,
you protect the environment and the health of your fellow human beings. The environment
and health are endangered by incorrect disposal. Material recycling helps to reduce the con-
sumption of raw materials. For further information on how to recycle this product, please
contact your local authority or municipal waste disposal companies.
The instrument does not produce any toxic or harmful gases or substances. During the
measurement, make sure that there are no toxic or harmful substances in the microtiter
plate.
5
Instructions for using the Absorbance 96
The Absorbance 96 is designed to carry out fast and sensitive absorbance measurements.
It measures the optical density (OD) of samples at defined wavelengths.
Absorption
Absorption refers to the amount of light absorbed by a medium. Absorption reduces trans-
mission. Transmission is the ratio of incident to transmitted light. Accordingly, the degree of
transmission is calculated as follows
T = (I/I0)
Optical Density
Optical density is a measure of the attenuation of light radiation after it has passed through
a medium. Optical density is the logarithmic quantity that describes the reciprocal of the
transmittance T:
OD = log(I0/I)
Optical density is the absorbance of the sample plus other attenuating effects such as scat-
tered light due to turbidity. It is therefore necessary to avoid scattered light in order to meas-
ure absorbance correctly.
6
Instructions for using the Absorbance 96
provided for further processing. The Absorbance 96 has 96 detection units, which allow
measurement without a scanning mechanism.
In the upper part of the measuring instrument there are 4 light sources with different wave-
lengths and 4 interference filters. The four signal lights on the top of the reader show which
measuring channel is being used for the current measurement.
The microtiter plate is inserted manually into the slot of the device. There are two raised
locking-edges at the bottom of the slot, as well as an arrow for visual inspection that be-
comes wholly visible only after the microtiter plate has been completely inserted.
Figure 1. Visual representation of the product from the front, top. 1. Signal lights. 2. Slot for
microtiter plate. 3. Raised locking-edges. 4. Arrow.
On the back of the Absorbance 96 is a USB plug for connecting the instrument to a computer
via the included Micro-USB cable. Furthermore, there is an eject button for ejecting the mi-
crotiter plate out of the slot.
Figure 2. Visual representation of the product from behind, top. 5. Eject button. 6. Micro-
USB plug.
7
Instructions for using the Absorbance 96
With the Absorbance 96, ANSI/SBS Standard 96-well microtiter plates can be read.
When using microtiter plates, always regard the specifications of the manufacturer of the
microtiter plates. Not all microtiter plates of a manufacturer are the same in design, materials,
or configuration. The temperature stability within the microtiter plate can depend on the
type of microtiter plate used.
The Absorbance 96 is an analytical instrument with very low power consumption. It is oper-
ated by a Micro-USB cable and the total power consumption is generally less than 2.5 watts.
8
Preparing the product for use
Unpacking
Remove the packaging material and carefully place the unit on a firm and level surface.
! Make sure, that the rubber feet are completely attached to the underside of the de-
vice.
The device is packed in a specially designed cardboard box. Keep the packaging material. If
you return the device for repair, the original packaging material and box must be used.
!! Protect the device from moisture and dust during prolonged storage.
! Do not store the device next to heat sources and protect it from direct sunlight.
Transportation
Before transportation, make sure that the device is disconnected from the computer and
that it is turned off. Remove the cable from the instrument and make sure that there is no
microtiter plate in the instrument. Depending on the transport distance, use the original
packaging material. Make sure that the new location meets the requirements described in
chapter 3.3.
! Always transport the product well packed to avoid possible transport damage.
When unpacking the device, please check that the following components are present:
• Absorbance 96
• Micro-USB cable
• Hex key
• Microfiber cloth
• USB Stick
9
Preparing the product for use
Due to its small size, the Absorbance 96 can be operated very well in various working envi-
ronments.
However, the following warnings must be regarded. Ignoring them may lead to a reduction
in the expected lifetime, damage to the device, or measurement errors:
!! The device should stand on a level surface, free from dust and vibrations.
!! Do not operate the device near heat sources or under direct sunlight.
To install the Absorbance 96 App, insert the USB stick provided with the Absorbance 96
Reader into the computer and follow the steps below:
I To install the proper drivers and software on the computer, the user must have ad-
ministrator rights, or obtain such permissions.
I For Windows PCs: The computer must have at least windows 7 or above. Windows
10 and later versions function through plug and play. If an older version is being
used, drivers may need to be installed. These are available on the USB stick pro-
vided with the device.
Uninstallation of software
The Absorbance 96 App can be uninstalled using the computer’s standard uninstall proce-
dure.
Updates
10
Operation
4 Operation
Before commissioning the Absorbance 96, you should carefully read and understand the en-
tire manual, so that you are familiar with the system.
The Absorbance 96 requires a connection to the computer for power supply, to start the
experiment, and for data transfer. Insert the Micro-USB cable into the Absorbance 96 and
the USB port of the computer. The Absorbance 96 starts automatically when connected to
a computer via the Micro-USB cable and the software automatically establishes a connec-
tion.
!! Always use the included Micro-USB cable for the connection to your computer.
I Connect the instrument directly to your computer and do not use an external USB
hub.
I The device may only be operated at a USB port USB 2/USB 3 with 5 VDC and a
maximum of 3 A of a certificated computer (with certification mark of an approved
testing laboratory). Use an adapter cable on USB 3.1 with type C plug connection,
which ensures the profile 1: 5 [email protected] A.
When the Absorbance 96 is switched on, the signal lights on the top of the instrument light
up. Each time the Instrument is switched on, an internal self-test is performed to ensure there
are no malfunctions.
If no connection between the instrument and the computer is established, install the driver
for the USB connection. You can find it in the supplied USB Stick. See chapter 3.4.
Select “New” from the menu bar. Your experiment conditions can now be defined.
In the first section of the Setup Page you can name your assay/protocol. For the protocol
you can additional add a description, assign a supplier for the assay kit, or choose a logo.
Workflow Parameters
Wavelengths
The signal wavelength is the wavelength (in nanometers) at which the principal measure-
ment will be run.
11
Operation
The reference wavelength is the wavelength (in nanometers) at which the reference is
measured. The values measured at the reference wavelength will be subtracted from the
values measured at the signal wavelength.
If no reference wavelength is desired, click the “None” button beside the reference wave-
length.
Readout Method
In an “Endpoint” measurement, one measurement cycle is performed. Here you can choose
between the different evaluation methods described below.
In a “Kinetic” measurement, you can set the parameters for multiple measurement cycles
taken over a given time span.
I For best results, acclimatize the connected reader and plate to the ambient temper-
ature for at least 1 h.
12
Operation
I With kinetic measurements, a signal drift of about 1 mOD per hour might be observed.
Evaluation
The evaluation section allows you to choose which evaluative method will be used for the
readout.
No Evaluation
No evaluation will be used. The raw OD values will later be shown in plate format.
Quantitative
• Upon selection of the Quantitative evaluation method, a second section titled “Fitting
Method” will appear in the evaluation field.
• Click the drop-down menu and select one of the options: point-to-point, linear re-
gression, 4-parameter or 5-parameter regression.
• The selected evaluation method will be used for the standard curve and the data
analysis.
Qualitative
Upon selection of qualitative evaluation, the plate layout window will change and a formula
for the cut-off definition will appear.
The formula setup for the cut-off definition covers multiple formula types. These are listed
below.
Example 1: Subtraction
To replace the minus function, the user must simply enter a negative value. For example, if
the cut-off is:
13
Operation
To define a range, you may use the “Range” field in the cut-off editor. This allows you to enter
a percentage that will correspond to values above and below a given cut-off value, within
which the experimental values will be deemed intermediate, above which positive, and below
which, negative.
For example:
Sample is considered positive if: SampleOD > (Mean Pos. + 10%)
Sample is considered negative if: SampleOD < (Mean Pos. -10%)
Sample is considered intermediate if: (Mean Pos. – 10%) < Sample OD < (SampleOD + 10%)
It may be the case that the cut-off value is known for a given experiment and need not be
calculated. In this case, the “0 OD” option may be selected for the calculation and the desired
value entered into the offset.
Then:
I The examples shown in this manual do not represent actual data, rather function to
clarify software features, and may not be referenced for experimental methods or
procedure.
Plate Layout
In the plate layout section, the microplate can be filled out as desired.
Each well may be defined individually using the buttons below the plate.
I The format of the plate layout and the corresponding buttons are defined by the
evaluation method. Quantitative and qualitative evaluations have different fields
and buttons.
14
Operation
I Once the conditions/parameters of a given experiment have been set, these set-
tings may be saved as a protocol. All parameters, such as wavelength and plate
layout, will be saved.
I Wells without values (concentration or sample ID) are shown with a ring instead of
solid colour.
Quantitative Evaluation
When the quantitative evaluation is selected, the window will be as shown in figure 5.
To define a well, click on the desired button below the plate. You may then click and drag, or
individually define single wells. The blue ring indicates which field is currently selected; all
fields will be defined starting with the selected number.
To enter a concentration or sample ID, type the value in beside the number and press ENTER
to move to the next field.
To enter a unit, enter the desired unit in the first field and press ENTER, or choose a prede-
fined unit form the drop-down menu. All fields will be automatically filled out according to the
first selection.
To define, for example, two duplicate standard rows, click and drag the desired number of
wells, release, then click and drag again in the next row.
15
Operation
To set a blank or to empty a given well, click on the “Blank” or “Empty” button respectively,
then click the desired well or drag and drop across the plate.
I When blank wells are selected, the values of these wells will be subtracted in the
readout.
Qualitative Evaluation
Upon selection of qualitative evaluation, the plate layout window will look like that shown in
figure 6. Define the plate according to the description above.
After selecting the checkbox “Use predefined Standard curve”, you cannot assign any new
standards.
16
Operation
File Types
Quality Extension
Protocol .byop
Assay .byoa
Export .csv/.pdf
Procedure walkthrough
1. Empty Slot
2. Initialization
Click “OK” to start the initialization. The initialization is carried out, which is indicated by a
short illumination of the respective LEDs.
17
Operation
3. Insert Plate
After successful initialization, you will be prompted to insert the microtiter plate. Pay atten-
tion to the alignment of the microtiter plate. For orientation, a pictogram on the top of the
Absorbance 96 indicates the correct alignment of the microtiter plate. (The pictogram is a
schematic representation. The actual shape of the microtiter plate, especially the position of
the rounded corner, may vary and differ from the illustration. Base your alignment on the A1
Well).
After insertion, confirm in the software that the microtiter plate has been inserted by clicking
OK. The measurement will subsequently be carried out, accompanied by a short illumination
of the respective signal lights.
In order to guarantee a correct measurement result, please pay attention to the following
warnings:
!! Make sure that the microtiter plate was inserted in the slot completely. In that case
the arrow at the bottom of the slot is visible completely.
!! The bottom of the microtiter plate must be dry before it is inserted into the instru-
ment. If the microtiter plate is wet at the bottom, dry it with a paper towel or similar.
!! Shocks of the Absorbance 96 and of the microtiter plate can cause an unwanted
signal. Therefore, neither the Absorbance 96 nor the microtiter plate should be
touched during measurement.
!! Note that the system depends on a clean and dust-free environment. Therefore, do
not wear powdered safety gloves.
!! Ensure that no direct sunlight or other strong light sources shine on the Absorb-
ance 96 and especially into its slot.
3. Read Plate
18
Operation
You may now remove the microtiter plate from the Absorbance 96. To do this, push the eject
button on the rear side of the device.
Plate Overview: Provides an overview of the different OD values in their respective wells.
Undefined wells are not shown. Click on a given well to see the OD, label and, if applicable,
concentration.
Graph Overview: A representation of the data will be shown on the graph. The curve will be
dependent on the data analysis method chosen and will be fitted accordingly. By choosing
the right regression method, the distance from the curve to each point is minimized. Using
the standard curve, the concentration of each sample is calculated. The equation below the
graph reflects the regression method applied and offers further data.
Tabular View: Provides an overview of the various samples, wells, ODs and calculations.
Parameter Description
19
Operation
20
Information required by the user
All signal lights flash simultaneously Error, software shows error message
Device is off
No signal lights light up Device is waiting for input
Device is defective
All signal lights light up briefly in sequence Self-test after connection to power
Signal light of the respective measuring
Initialization/measurement in progress
channel flashes/illuminates continuously
If there is an error, all four signal lights flash simultaneously. In this case, check the software
to see which error is present.
In the event of the following errors, a new measurement can be carried out immediately after
the error has been corrected.
Restart
Type of error Cause Solution
necessary?
There was a microtiter
Remove the plate/all
plate in the slot during No
objects
initialization.
Initialization failed Clean the inside of the
The inside of the slot slot according to the
No
is dirty cleaning instructions
in chapter 6.2.
Make sure the instru-
ment is not exposed to
Interference due to Too much ambient
too much ambient
ambient light that is light is entering the No
light, e.g. direct sun-
too bright slot
light, during the meas-
urement process.
The ambient temper- Make sure the ambient
Temperature out of
ature was too high or temperature is be- No
specification
too low tween 5-45°C
Use a different USB
USB Power Supply Yes. Remove
port on the computer
Defective (<450mA or Defective USB port cable and re-
or a different com-
< 4V) connect it.
puter
21
Information required by the user
22
Maintenance and cleaning
6.1 Maintenance
The Absorbance 96 is maintenance-free. Each time the instrument is switched on, an internal
self-test is carried out to ensure there are no malfunctions.
There are no parts within the ABSORBANCE 96 that can be serviced by the customer. It is
only necessary to ensure that the device is kept clean.
The accuracy of the device can be checked with a reference plate. If you check more than
one point for accuracy, you can also confirm the linearity of the reader. Byonoy recommends
the reference plate from Hellma GmbH (item code 666-R013), for which the following steps
are described:
1) Measure the plate at the desired wavelength. Ensure that there is no dust or other
contamination on the reference plate.
ODRP: Optical density of the reference plate at the desired wavelength. This value is
found on the calibration certificate from the manufacturer.
MU: Measurement uncertainty of the reference plate. This value is found on the cali-
bration certificate from the manufacturer.
SP: Accuracy specification of the reader at measured optical density. This value is
found in chapter 7.
ODRES: The optical density measured by the reader at the desired wavelength.
Example:
ODRP: 1.554
MU: ± 0.0079
ODRES: 1.573
The measured Optical Density of the reader lies within the calculated range and
hence the accuracy of the reader is verified.
4) If all Results (ODRES) are within the range, the accuracy and linearity of the device
have been verified.
23
Maintenance and cleaning
In case that the results are outside the range, please contact the technical service or the
local responsible person.
I Please follow the plate manufacturer’s recalibration instructions for the reference
plate.
6.2 Cleaning
!! Make sure before cleaning, that there is no microtiter plate in the instrument.
The surfaces of the housing should be cleaned regularly. For this purpose, you can use e.g.
a cloth, or a sponge lightly wetted in water. For heavier soiling, clean the surface of the hous-
ing with a mild soap solution diluted with water or glass cleaner and then wipe with a lightly
moistened cloth or sponge to remove any residue. Do not use scouring agents.
To clean the slot of the RIDA ABSORBANCE 96, first open the four screws on the bottom of
the instrument (see Figure 11) using the Hex key provided.
I Do not loosen any screws that are not explicitly marked in the manual, as this can
lead to a malfunction and may void the instrument's warranty.
24
Maintenance and cleaning
Figure 10. Pull the instrument apart into its two components
Carefully pull apart the top and the bottom of the instrument to unplug the plug connection
on the back of the instrument.
I To avoid damaging the rigid plug connection between the top and bottom of the in-
strument, grasp the instrument at the back during the pull movement. Avoid any
tilting movement.
Use a dry microfiber cloth or a bellows to carefully remove dust or dirt from the surface. If
this is not enough, the microfiber cloth can be slightly soaked with 80% ethanol to carefully
remove dust or contamination from the surface.
Do not allow water or other liquids to get inside the instrument. If this happens, return the
device to the manufacturer for inspection.
When cleaning, pay attention to the following warnings. The non-observance of these warn-
ings may result in damage to the user or to the instrument. This can lead to a reduction of
the service life or in measurement errors:
!! Always avoid spraying liquid directly onto the surfaces of the ABSORBANCE 96.
This is especially important for the inside of the slot, where there are optical ele-
ments that are very sensitive and essential for the functioning of the instrument.
!! Never clean the inside of the slot with sharp, abrasive, solvent-containing agents.
!! Biological hazard: Always wear gloves during cleaning operations that may be asso-
ciated with contact with biological or generally hazardous materials or liquids.
25
Maintenance and cleaning
In case of problems with the instrument, please contact the technical service or the local
responsible person.
For decontamination of the Absorbance 96, follow the government guidelines for inactiva-
tion of organisms used in biological laboratories.
6.4 Repairs
Repairs on the device may only be carried out by the manufacturer. Please contact the ser-
vice department. The product liability expires if the device is modified by unauthorized per-
sons and by the installation of parts of a different type.
26
Performance data and technical data
Parameter Value
Type of product Absorbance reader
Product name Absorbance 96
Service life 10 years at an average use of 4h/day
Housing material Aluminium
Place for use Laboratory
Degree of contamination 2
Temperature for storage/measurement 5-45°C
Temperature (Transport) -10-50°C
Relative tolerated humidity (storage/measure-
0-70%
ment)
Air pressure (measurement) 900-1070 hPa
Air pressure (storage/transport) 600-1070 hPa
Elevation Max 5100m
Measurement method Absorbance
Measuring technique Endpoint and Kinetic
Microplate types 96-well, fat shaped bottom
Light source 4 LEDs
Filters 4
Detection 96 Photodiodes
Measurement range 0-4.0 OD
Resolution 0.001 OD
0,000–2,000 OD ≤ (1 % + 0,010 OD)
Accuracy*
2,000–3,000 OD ≤ (1,5 % + 0,010 OD)
0,000–2,000 OD ≤ (0,5 % + 0,005 OD)
Reproducibility**
2,000–3,000 OD ≤ (1 % + 0,010 OD)
0,000–2,000 OD ≤ 1 %
Linearity***
2,000–3,000 OD ≤ 1,5 %
Connection to computer USB 2/USB 3 with 5 VDC and max. 3A
Dimensions 55 x 96 x 154 mm
Power supply 5 VDC
Nominal value/Characteristic of the fuse 1A/very fast-acting
Power input 2.5W
Weight 900g
*Accuracy is the maximum deviation between the determined value and the true value;
**Reproducibility is the maximum deviation between the determined values when the
measurement is repeated directly;
***Linearity is the maximum deviation between the true and the determined increase of
the value.
27
Guarantee
8 Guarantee
The warranty period of the Absorbance 96 is 24 months. If a defect occurs in your device
during the warranty period, please contact our service staff directly. The instrument may
only be operated in technically perfect condition.
In the event of defects that could endanger employees or third parties, the device may only
be used again after it has been repaired. This warranty does not cover damage caused by
improper use or external mechanical influences, transport damage or unauthorized inter-
vention in the device by unauthorized persons.
28
EC-Conformity Declaration
EG-Konformitätserklärung
EC-Conformity Declaration
The Product named below fulfills the relevant fundamental requirements of the EC direc-
tives and standards listed. In the case of unauthorized modifications to the product or an
unintended use this declaration becomes invalid.
Produkttyp
Product type
Absorptionsmessgerät
Absorbance reader
Typenbezeichnung
Type designation
Absorbance 96
Einschlägige EG-Richtlinien/Normen
Relevant EC directives/standards
2011/65/EU; 2014/30/EU;
DIN EN ISO 14971:2013-04; DIN EN 62366-1:2017-07; DIN EN 62304:2016-10; DIN EN
61010-1:2011-07; DIN EN 61010-2-101:2017-10; DIN EN 61326-1:2013-07; DIN EN 61326-2-
6:2013-07
29