DNA Extraction Protocol - Watch a Video

DNA extraction is crucial for acquiring an initial sample for techniques such as PCR, DNA sequencing, and electrophoresis. However, these processes require a high yield and the removal of impurities (such as cell debris) to successfully provide truthful findings. For example, the presence of contaminants such as DNases can lead to the breakdown of DNA and so reduce the overall yield, potentially below the required sample amount.

This video will help optimize your DNA extraction technique, clearly explaining and offering tips on how to achieve efficient, reliable results; avoiding contamination and poor yield.

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Protocol 1 DNA Extraction

Source: Youtube via  The Jackson Laboratory

A path to the perfect pipetting

Understanding Pipettes

Pipette terms:

• Adjustment - altering the pipette so that the dispensed volume is within the specifications.
• Air Displacement Pipettes - are meant for general use with aqueous solutions. In air displacement pipettes, a certain volume of air remains between the piston and the liquid.
• Aspirate - to draw up the sample.
• Blow-out - to empty the tip completely.
• Calibration check - checking the difference between the dispensed volume and the selected volume.
• Dispense - to deliver the sample.
• Positive Displacement Pipettes - are used for high viscosity and volatile liquids. In positive displacement pipettes, the piston is in direct contact with the liquid.

The effect of the pipetting position (image)

1. Pipette vertical, tip immersed about 1 cm into the liquid.
2. Pipette vertical, tip immersed about 3 cm into the liquid.
3. Pipette at a 30 - 40 degree angle; tip immersed about 3-4 cm into the liquid.

Find more useful hints here:
Pipettes, Calibration & Repair Service

Source: Pipette.com

The good and bad of microbiology lab automation

Here is a summary of the potential quality benefits of “Total Lab Automation” (TLA) systems (e.g. Kiestra, Biomeurieux, Copan Wasp). You might be involved in a business case for one for your own laboratory.

Main advantages:

• Plate tracking
• Less menial tasks
• Better plate spreading
• Less plate contamination
• Storage of digital images of plates
• Standardised incubation times
• Less time out of the incubator
• Remote plate reading
• Plate checking
• Plate interpretation

Disadvantages:

• Redundancy of staff
• What happens if it breaks down<
  

Read more:
“The good and bad of lab automation….

Source: Microbiology matters
Image: Michael

MALDI-ToF - Benefits and Limitations in Microbiology

Helping clinicians quickly and effectively address the medical concerns of their patients should be the ultimate goal of any clinical microbiology laboratory. Delivering on this goal can be especially important when a patient presents with a bacterial or fungal infection. MALDI-ToF can help improve patient outcomes by speeding diagnosis and improving the quality of care.

There are many benefits to incorporating MALDI-ToF to aid in identification of microorganisms. Identification with MALDI-ToF is a relatively simple process. The process can reduce the time needed for identification and diagnosis from days to hours. There are, however, some limitations to MALDI-ToF. Some organisms may require repeat analysis and additional processing, and some closely related organisms are not differentiated and may cause challenges with identification. Also the initial purchase price of the equipment is high, which may limit the types of labs that can adopt it.

While not all laboratories are capable of widely adopting MALDI-ToF, those that can, probably should. MALDI-ToF may promote better patient outcomes through speedier diagnosis and information to influence efficacious treatment for many bacterial, yeast, and mycobacterial infections, greatly improving the quality of clinical management and patient care.

Read more:

MALDI-ToF is poised to speed diagnosis for bacterial and fungal infections(MLO)

MALDI-ToF principle (Frontiers Microbiology)

Source: MLO
Image: Juha Wahlstedt

This USB Stick can perform an HIV Test

Scientists in the UK have developed a USB stick that can quickly and accurately measure the amount of HIV is in a patient’s blood.

The medical device was created by scientists at Imperial College London and tech firm DNA Electronics, and all it needs is a simple drop of blood to measure HIV-1 levels. It then creates an electrical signal that’s fed into a computer, laptop, or handheld device. The disposable test could be used by HIV patients to monitor their own treatment and help patients in remote regions of the world, where more standard HIV tests are inaccessible.

The timely detection of viremia in HIV-infected patients receiving antiviral treatment is key to ensuring effective therapy and preventing the emergence of drug resistance. In high HIV burden settings, the cost and complexity of diagnostics limit their availability. Scientists in the UK have developed a novel complementary metal-oxide semiconductor (CMOS) chip based, pH-mediated, point-of-care HIV-1 viral load monitoring assay that simultaneously amplifies and detects HIV-1 RNA. A novel low-buffer HIV-1 pH-LAMP (loop-mediated isothermal amplification) assay was optimised and incorporated into a pH sensitive CMOS chip. Screening of 991 clinical samples (164 on the chip) yielded a sensitivity of 95% (in vitro) and 88.8% (on-chip) at >1000 RNA copies/reaction across a broad spectrum of HIV-1 viral clades. Median time to detection was 20.8 minutes in samples with >1000 copies RNA. The sensitivity, specificity and reproducibility are close to that required to produce a point-of-care device which would be of benefit in resource poor regions, and could be performed on an USB stick or similar low power device.

Read more:
Novel pH sensing semiconductor for point-of-care detection of HIV-1 viremia

Source: Nature: Scientific reports
Image: Imperial College London/DNA Electronics

Lab Cam

iPhones take great pictures. This adapter makes it super easy to take images and make videos with your iPhone through your microscope. You can even use your iPhone to live project/stream your view. The iDu adapter fits iPhone6/6s. It's fitted with a 10x magnifying lens and comes with two adapters to fit a 30 mm or 23 mm eyepiece slot (it should fit all Nikon, Olympus, Zeiss, Leica and other common brand microscopes).

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iDu Optics LabCam Microscope Adapter for iPhone 


Source: Youtube

Microscope Koehler Illumination Online Guide

In almost all advanced imaging techniques there is a need for a correct transmitted light illumination. This tutorial explains the correct setup for Koehler illumination based on a motorized high-end inverted microscope used for transmitted light techniques (phase contrast, DIC etc.) and fluorescence microscopy. Explore the cross-sectional models of the optical pathway. Learn more about relevant components for Koehler Illumination.

Step by Step Guide to Optimal Illumination of Specimen

Open the guide here:
Koehler Illumination

Source: Leica Science Lab

Bike-Powered Centrifuge

Designer Jack Albert Trew, a 3D Product Design student at the University of Birmingham City, has designed a low-tech centrifuge alternative to diagnose medical patients in rural developing countries.

Most often, Western medical devices donated to Africa have limited shelf life due to electrical needs and lack of budgets for constant care and repairs.

The World Health Organization estimates that a large proportion—up to 70 percent—of donated medical equipment lies idle. This leads to an inadequacy of donating medical devices to Africa, and a waste in equipment that can potentially save many lives.

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Bike-Powered Centrifuge for Developing CountriesSource

(Click image to enlarge)

Source: BORGEN

12 must-read tips to improve your pipetting technique

When was the last time you thought about your pipetting technique after you first learned about it? For many of us pipetting becomes routine after a little practice. Typically, liquid handling quality assurance regulations place great emphasis on pipette calibration, repair and maintenance. However, ensuring the competence of the pipette operator (a lab employee) is an often neglected and altogether crucial area. In reality even highly experienced laboratory technologists may have never received formal pipetting training and may be prone to routine errors.

Read more:
Pipetting Skills Test – How Good Are You? 

Source: Splice

New Point-of-Care Test For Male Fertility Potential

More than 70 million couples worldwide are affected by infertility, with male-factor infertility accounting for about half of the cases. Semen analysis is critical for determining male fertility potential, but conventional testing is costly and complex. Here, we demonstrate a paper-based microfluidic approach to quantify male fertility potential, simultaneously measuring 3 critical semen parameters in 10 min: live and motile sperm concentrations and sperm motility.

Detection limits of 8.46 and 15.18 million/mL were achieved for live and motile sperm concentrations, respectively. The live and motile sperm concentrations and motility values from our device correlated with those of the standard clinical approaches (R2 ≥ 0.84). In all cases, device provided 100% agreement in terms of clinical outcome. The device was also robust and could tolerate conditions of high absolute humidity (22.8 g/m3) up to 16 weeks when packaged with desiccant.

New device outperforms existing commercial paper-based assays by quantitatively measuring live and motile sperm concentrations and motility, in only 10 min. This approach is applicable to current clinical practices as well as self-diagnostic applications.

Read more:
Paper-Based Quantification of Male Fertility Potential

Sinton Lab web page 

Source: Clinical Chemistry



Source: Clinical Chemistry & Sinton Lab

T2Candida Panel - A Blood Scan for Sepsis Without Culture

The T2Candida panel and the T2Dx Instrument are for the direct detection of Candida species in human whole blood specimens from patients with symptoms of invasive fungal infections or with conditions that predispose them to such infections.

The T2Candida panel detects Candida albicans, Candida tropicalis, Candida parapsilosis, Candida glabrata, and Candida krusei. The FDA said that the panel is the first direct blood test for detecting the five yeast pathogens.

According to the agency, traditional methods of detecting yeast pathogens in the bloodstream can take up to six days and identifying the specific type of yeast present in the bloodstream may take even longer. By comparison, the T2Candida panel and T2Dx Instrument can identify the five common yeast pathogens from a single blood specimen within three to five hours.

"By testing one blood sample for five yeast pathogens — and getting results within a few hours — physicians can initiate appropriate anti-fungal treatment earlier, and potentially reduce patient illness and decrease the risk of dying from these infections,"

Read more
FDA Clears T2 Bio Test, System for Identification of Yeast Pathogens'

T2Biosystems web page 

Source: Genome web

Blood Cell Flower

Beautiful 3D science animations.

Illustrations, 3d animations and multimedia about medical, bitechnology, mechanisms of action and popular science. Do not matters the complexity of ideas, graphic concepts to convey or its scale, it can be visible and understandable to a much wider audience. Furthermore, this can be achieved joining visual impact and aesthetics with accuracy and clearly. At most cases than this requires decipt the invisible or the inaccessible.

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3dciencia

 Source: 3Dciencia

New Microfluidic Blood-draw Device Could Replace Needle Sticks

Tasso is revolutionizing diagnostics by creating technologies for blood sample collection that place the user at the center of process. Our wearable blood collection system, the HemoLinkTM, is being developed to improve the user experience as well as enable both healthcare consumers and providers with convenient access to reliable laboratory test results.

In a process known as “capillary action,” HemoLink leverages microfluidics to create a slight vacuum that pulls blood from capillaries though tiny channels in the skin into a small tube, noted a Gizmag report. The device collects 0.15 cubic centimeters of blood, which is enough to test for cholesterol, infections, cancer cells, blood sugar and other conditions.

Pathologists and clinical laboratory professionals will be watching the eventual launch of HemoLink to learn how its developers have overcome the problems affecting lab test accuracy that can be caused by the interstitial fluid that often accompanies capillary blood when such specimens are collected. How the lab test technology used by Theranos addresses this same problem has been an ongoing point of interest among medical laboratory professionals.

Read more:
New Microfluidic Blood-draw Device Could Replace Needle Sticks and Venipunctures at Medical Laboratories

Source: Dark Daily

Dissolvable electronics monitor health inside the body and then melts away

From the folds and crinkles of a living brain, a fleeting fleck of electronics smaller than a grain of rice can wirelessly relay critical health information and then gently fade away.

The transient sensors, which can measure pressure, temperature, pH, motion, flow, and potentially specific biomolecules, stand to permanently improve patient care, researchers said. With a wireless, dissolving sensor, doctors could ditch the old versions that require tethering patients to medical equipment and performing invasive surgery to remove, which adds risks of infections and complications to already vulnerable patients.

The devices can also be customized with different types of sensors and coatings. The thicker the coating, the longer they take to dissolve, Murphy explained. The current versions can last for a few days in biofluids, including cerebrospinal fluid, at physiological temperature. But, Murphy said, the researchers are hoping to make versions that can last for weeks.

Read more:
In a brain, dissolvable electronics monitor health and then vanish 

Source: Ars Technica

Doctors 3D-print living body parts

Custom-made, living body parts have been 3D-printed in a significant advance for regenerative medicine, say scientists.

The Integrated Tissue and Organ Printing System - or Itop - combines a bio-degradeable plastic which gives the structure and a water-based gel which contains the cells and encourages them to grow. When the structures were implanted into animals, the plastic broke down as it was replaced by a natural, structural "matrix" of proteins produced by the cells. Meanwhile, blood vessels and nerves grew into the implants. Prof Anthony Atala, the lead researcher, said tissues could now be printed on a human scale.

The breakthrough, published in Nature Biotechnology, raises the hope of using living tissues to repair the body.

Read more:
Doctors 3D-print 'living' body parts

Source: BBC News

Super-resolution microscopy as a potential approach to platelet granule disorder diagnosis

Many platelet functions are dependent on bioactive molecules released from their granules. Deficiencies of these granules in number, shape or content are associated with bleeding. The small size of these granules is such that imaging them for diagnosis has traditionally required electron microscopy. However, recently developed super-resolution microscopes provide sufficient spatial resolution to effectively image platelet granules. When combined with automated image analysis, these methods provide a quantitative, unbiased, rapidly acquired dataset that can readily and reliably reveal differences in platelet granules between individuals.

A super-resolution imaging approach is effective and rapid in objectively differentiating between patients with a platelet bleeding disorder and healthy volunteers. CD63 is a useful marker for predicting Hermansky-Pudlak Syndrome and could be used in the diagnosis of patients suspected of other Platelet granule disorders.

Read more:
Super-resolution microscopy as a potential approach to platelet granule disorder diagnosis - Westmoreland - Journal of Thrombosis and Haemostasis 

Source: JTH Journal of Thrombosis and Haemostasis

A brief history of microscopy

History of microscopy from 2000 BC till present.

Read more:
A brief history of microscopy

Source: I heart histo

A new biosensor test system based on magnetic nanoparticles suitable for POCT

Researchers from the General Physics Institute of the Russian Academy of Sciences (GPI RAS) and Moscow Institute of Physics and Technology (MIPT) have developed a new biosensor test system based on magnetic nanoparticles. It is designed to provide highly accurate measurements of the concentration of protein molecules (e.g. markers, which indicate the onset or development of a disease) in various samples, including opaque solutions or strongly coloured liquids.

At molecular level, the magnetic nanoparticles "link" with antibodies to the required protein and then they are placed on a porous plate close to the intended point of contact with the test solution. The liquid, which spreads along the plate due to capillary action, captures the magnetic particles. It then meets two lines - the test line and the control line. The test line contains antibodies that capture the protein in question and also the magnetic markers that became attached to molecules of the protein due to the fact that the nanoparticles are also "linked" to the antibodies. The control line only captures the antibodies with magnetic markers, and it will be activated in any case, if the test strip is usable. The control line serves as an indicator as to whether the test is suitable for use, the protein antibodies in it have not been destroyed due to improper storage, and the test liquid has been applied correctly.

The combination of reliability, accessibility, and high accuracy and sensitivity of the new method means that it is likely to make a rapid transition from a laboratory prototype to mass production.

Read more:
Scientists have put a high precision blood assay into a simple test strip

Source: EurekAlert! Science News

Lumify Mobile Ultrasound Imaging

With Lumify, high-quality ultrasound is available almost anywhere. Just subscribe, download the Lumify app, plug in the transducer, and you're set. Meet patients at the point-of-care, get a faster diagnosis, and deliver care whenever it's needed.

Read more:
Lumify | Ultrasound Imaging

Source: Phillips

Improving Real-Time PCR Data Quality

The polymerase chain reaction (PCR) has radically transformed biological science, allowing sophisticated analysis of genes and the genome. Revolutionizing the study of DNA, PCR is often hailed as one of the most important scientific advances of the 20th century. Over time, PCR has evolved into fluorescence-based quantitative real-time PCR (qPCR), which is now considered the molecular diagnostic technique of choice due to its capacity to detect and measure minute amounts of nucleic acids in a variety of samples from multiple sources.

Due to its practical simplicity, in combination with its outstanding capabilities, including speed, sensitivity, and specificity, qPCR plays a huge role in a number of applications, among them gene expression analysis, microRNA analysis, single nucleotide polymorphism genotyping, copy number variation analysis, and protein analysis.

The success and reputation of qPCR is reflected in the abundance of publications reporting qPCR data. Despite substantial advances in the accessibility and ease-of-use of qPCR for diagnostics, generating an assay that is capable of delivering reliable, reproducible, and meaningful results is still a challenging task.

Read more:
Improving Real-Time PCR Data Quality

Source: GEN Magazine Articles