Wednesday, August 16, 2017

NADH Analysis Just Got a Lot Easier!

The next generation 800™ TS Absorbance Reader was recently introduced in a previous blog posting. The many enhancements to both hardware and software were designed to simplify workflows for many of the most commonly performed assays. Assays such as ELISA, UV-absorption and colorimetric absorbance are performed in a variety of laboratory settings ranging from basic research in academic labs to clinical laboratory testing.

A long studied enzyme family found in a diverse range of organisms includes a group of alcohol dehydrogenase enzymes (ADH) which are responsible for interconversion between alcohols and aldehydes or ketones. The enzymatic catalysis of alcohol requires the reduction of the cofactor nicotinamide adenine dinucleotide (NAD+) to the reduced form NADH or the reverse chemical process for the formation of alcohol.

NADH

The spectral properties of the reduced form NADH, show an absorption peak at 340 nm providing a simple method to monitor progression of a reaction.  The enzymatic activity of ADH from S. cerevisiae was determined for the two alcohols, ethanol and 2-propanol, by obtaining kinetic absorbance measurements at 340 nm. Reaction velocities were calculated using Gen5™ Data Analysis software; Michaelis-Menten and Lineweaver Burk plots were generated to determine K­m and Vmax for each reaction.


These and similar experiments are made simple by the capabilities of the new, easy-to-use 800 TS. The 800 TS brings excellent performance, high quality and value to many laboratories. It's no wonder that BioTek continues to be the most trusted and well-known name in microplate readers and other life science instrumentation.


By: BioTek Instruments, Peter J. Brescia Jr., MSc, MBA

Tuesday, August 8, 2017

BioTek Unveils Next-Gen Plate Reader and Washer

In 1995, I started my career with BioTek. I had just graduated with a degree in a Clinical Laboratory Science and was hired as a Service Engineer in BioTek's Technical Assistance Center. The job's primary focus was helping customers program, utilize and maintain their microplate readers and washers. Since I was trained as a Medical Technologist, I had a fair amount of experience with ELISA instrumentation. I had actually used a BioTek EL312 plate reader in my clinical rotations. I recall being very impressed by the automation and speed the reader gave us with its ability to process several samples at a time, compared to my experience with a manual cuvette spectrophotometer.

The year I started working at BioTek, we launched the ELx800 Microplate Absorbance Reader and shortly thereafter, the ELx50 Automated Strip Washer. This pair of workhorses had been entirely re-imagined from predecessors like my EL312 back in school. With sleek hardware profiles, powerful data reduction and easy to use interfaces, these products were cutting-edge. As the years went by and I transitioned to Product Management, I always found the variety and increasing number of assays customers ran on the instruments fascinating – from human and veterinary diagnostics to food safety testing and all areas of research.

Now more than 20 years later and nearly 50,000 installations around the world, BioTek has once again unveiled a next generation. The new 800 TS Absorbance Reader and 50 TS Washer have been redesigned for today’s busy labs and individualized workflows, complete with color touch screens and USB flash drives for convenient data export. Just like the ELx800 and ELx50, the new instruments offer high reliability, excellent performance and ease-of-use – all at an affordable price! After almost 50 years, BioTek products are still manufactured in Vermont, USA to the highest standards. I was lucky to have found this employer many years ago and am proud of the high quality products we continue to produce.

800 TS Absorbance Reader and 50 TS Washer
800 TS Absorbance Reader and 50 TS Washer


To celebrate the 800 TS and 50 TS launch, BioTek has just announced a giveaway contest for our thousands of loyal customers around the world. Do you have an ELx800 or ELx50 in your lab? If so, we want to thank you for making us the most trusted name in plate readers and washers. We would love to hear about how you use these instruments and how long they have been on your bench. All you need to do is visit www.biotek.com/contest and tell us your story by October 31, 2017. You could win a brand new 800 TS and 50 TS for your lab!

Microplate Reader and Washer Giveaway



By, BioTek Instruments, Jason Greene, Senior Product Marketing Manager

Tuesday, July 18, 2017

Performance of Image-Based Label-Free Tracking and Quantification of T-Cell Activation for Use in Cancer Immunotherapy Applications


Interest in cancer immunotherapy research has dramatically increased in recent years, due to the success of a variety of therapies now available, including:
  • cancer vaccines
  • antibodies or other proteins used to either stimulate or block immune checkpoint pathways
  • cells from a patient's own immune system (natural killer or T-cells)
One particular area of interest is the use of activated T-cells to target particular cancers. This has led to unprecedented results where 94% of patients with acute lymphoblastic leukemia (ALL) saw symptoms completely disappear in one study and > 80% of blood cancer patients experienced remission in a second study1. The process involves removing a portion of the patient’s peripheral blood, isolating the T-cells, activating and expanding the isolated cells, and finally reintroducing them back into the patient2.

A critical part of the process is the directed activation and expansion of isolated T-cells. Following antigen binding, as the cells become activated, increased proliferative rates are exhibited and are visualized as aggregates in a clustering effect within the well3. To properly track and quantify these phenotypic changes, cell imaging is essential. The brightfield channel of BioTek’s Cytation Cell Imaging Multi-Mode Reader allows for label-free monitoring of T-cell activation. When integratedas part of the BioSpa Live Cell Imaging System, the results from multi-day activation experiments can be easily quantified.

In the experiment below isolated T-cells were placed into wells of a 24-well plate at a concentration of 100,000 cells per well in the presence of anti-CD3 and anti-CD28 antibodies, plus varying concentrations of an IL-2 Superkine (AdipoGen Life Sciences).

video
250 ng/mL anti-CD3/250 ng/mL anti-CD28/ 100 ng/mL IL-2 Superkine

video
3MB - 250 ng/mL anti-CD3/250 ng/mL anti-CD28/ 0 ng/mL IL-2 Superkine

Higher levels of cell proliferation, as indicated by an increase in dark, highly confluent areas within the image, are observed when the cells are in the presence of the antibodies and IL-2 superkine, compared to antibodies alone.

Quantification of activation can also be achieved through determination of the level of confluency and area covered by cell aggregates.

T-Cell Activation


The results validate the ability of BioTek’s instrumentation to be used for this critical step of the T-cell immunotherapy process.

  1. Yuhas, A. Cancer researchers claim 'extraordinary results' using T-cell therapy. The Guardian, [Online] Feb 15, 2016. https://www.theguardian.com/science/2016/feb/15/cancer-extraordinary-results-t-cell-therapy-research-clinical-trials (accessed Jul 14, 2017). 
  2. Topalian, S.L.; Weiner, G.J.; Pardoll, D.M. Cancer immunotherapy comes of age. J Clin Oncol. 2011, 29, 4828–4836. 
  3. Purtic, B.; Pitcher, L.A.; van Oers, N.S.C.; W├╝lfing, C. T cell receptor (TCR) clustering in the immunological synapse integrates TCR and costimulatory signaling in selected T cells. PNAS. 2005, 102(8), 2904-2909. 

By: BioTek Instruments, Brad Larson, Principal Scientist

Tuesday, June 20, 2017

Zebrafish Development: From a Few Cells to an Embryo

Zebrafish are a great model organism used to study numerous biological processes.  Some of the benefits of the zebrafish model are that they mature within a short period of time, they are optically transparent, develop outside of the mother’s body, and are vertebrates.  Because of these, and other reasons, zebrafish are used in a wide variety of applications including drug discovery, developmental biology, and molecular genetics. Zebrafish are easy to analyze in early development, as the embryo is relatively easy to manipulate and visualize.

video

Zebrafish embryonic development: from 16 cells to 1 day

The video above is a great example of this.  In the video, the development of a zebrafish embryo is highlighted, progressing from a few cells to a 1 day old embryo where the heart is beginning to beat and the blood is beginning to flow. This video was provided from a Cytation 5, with images taken every 2 minutes for 24 hours. The very first panel of the video shows an embryo with about 16 cells sitting on top of the yolk sac.  The embryo is surrounded by a protective membrane called a chorion.  For the first week of its life, the embryo will get all the nutrients it needs from the yolk sac. After the initial division, the cells of an embryo divide every 15 minutes.  Therefore, the cell number quickly progresses from 16 to 128 in one hour.  The cell mass forms a ball of cells on the yolk sac and this point marks the beginning of the blastula period.  During this part of development, the cells change from a high piled cell mound to a cup-shaped multilayer. This multilayer of cells begins to thin and spread over the yolk sac, and the embryo changes shape from spherical to oblong.

16 cell zebrafish embryo
16 cell zebrafish embryo
The next stage of development is gastrulation. Gastrulation starts at 50% epiboly, or when the cell layer is covering about half of the yolk sac. This is the stage when the primary germ layers (endoderm, mesoderm, and ectoderm) are formed. The gastrula period is over when epiboly is complete (i.e. when the cell layer is covering the whole yolk sac) and the tail bud has formed marking the posterior of the embryo. 
Zebrafish embryo during segmentation period
Zebrafish embryo during segmentation period
The next portion of development is my favorite; it is known as the segmentation period. This is when the embryo starts to look like something more than a bundle of cells and the eye and somites begin to appear.  Somites are blocks of mesoderm that form along the anterior-posterior axis of the embryo that will eventually give rise to skeletal muscle, vertebrae, and skin.  Halfway through the video you can see the eye in the upper right hand corner of the embryo, the tail in the bottom right hand side of the embryos, and the somites forming along the body. After the somites appear, you will notice that the embryo body begins to lengthen and eventually the embryo begins to move for the first time.  By the end of the video pigmentation has started to develop on the eye and in the skin and the embryo is at the beginning of the pharyngula period. The heart is just beginning to beat, the blood is beginning to circulate and the embryo is quite active. In just 24 hours the zebrafish embryo goes from a few cells to a live, moving, organism!

For more information check out:



By: BioTek Instruments, Sarah Beckman, PhD., Microscopist
 

Thursday, March 30, 2017

BioTek at AACR 2017

The American Association for Cancer Research (AACR) will hold its annual meeting next week from April 1-5 in Washington D.C. The meeting will celebrate an important anniversary and marks a homecoming of sorts as the association was founded 110 years ago in Washington D.C. This conference represents the largest gathering of cancer researchers and clinicians from around the world, as each year more than 18,000 people attend to learn about new advances within oncology.

One area promising to receive a great deal of interest this year is the field of immuno-oncology. Here the goal is to stimulate a patient’s own immune system to fight the specific cancer. Therefore, new drugs, or immunotherapies, are developed to target not the individual cancer cells, but particular cells of the immune system, such as natural killer (NK) or cytotoxic T lymphocytes (CTLs). BioTek will present an Exhibitor Spotlight Tutorial in cooperation with Lonza and Greiner Bio-One entitled “Demonstration of High Throughput, Image-Based 2D and 3D Natural Killer Cell Mediated Cytotoxicity Assays” to illustrate how in vitro immunotherapy optimization experiments can be performed in a walk-away manner and analyzed using highly sensitive cellular imaging. The tutorial will take place on Tuesday, April 4th from 4-5:00pm in Spotlight Theater B. We invite you to attend the talk and to stop by our booth (1857) to learn about all the 2D and 3D oncology applications optimized using BioTek’s instrumentation.

3D HCT116 tumoroid exhibiting apoptosis and necrosis following NK cell exposure.
3D HCT116 tumoroid exhibiting apoptosis and necrosis following NK cell exposure.
 
By: BioTek Instruments, Brad Larson, Principal Scientist

Thursday, March 2, 2017

Free up some time: 2017 SLAS

This was my first year attending the SLAS meeting. I was impressed with the tremendous breadth of automated instrumentation available to life science professionals. Being a bit of a gear-head, it was easy being captivated by all the moving arms, spinning pumps, traveling vessels and liquids being moved about. At the end of the day though, it really comes down to determining the bottlenecks in a particular workflow and how this myriad of technologies helps free up more time to manage other tasks.
BioTek Instrumentation at SLAS

Spending time listening to other scientists and automation partners visiting the BioTek booth provided excellent examples of how BioTek instruments are currently being used and ideas for future use as both standalone workstations or by incorporating into larger automated work cells. Examples ranged from an automated washing and dispensing workstation for up to 100 plates per day to live cell experiments lasting weeks by incorporating an EL406 Washer Dispenser and BioSpa Live Cell Imaging System to manage media exchanges and kinetic imaging/plate reading.

It appeared that once again the BioSpa was a big hit drawing the attention of many passers-by. Folks were pleased to find an automated solution with such a small footprint, intuitive control and scheduling software and flexibility to interface with a variety of instruments. Of equal interest was the Lionheart FX Automated Live Cell Imager for the same small footprint and integrated environmental control for longer term kinetic experiments on cells.

BioTek booth
 
As a testament to the practical use of the BioSpa System the Scientists’ Choice Award for Best Drug Discovery & Development Article included the use of the system in combination with an EL406 for research into cancer-causing genetic abnormalities and potential therapies.

Customers at BioTek booth at the SLAS
 
The meeting was a fantastic opportunity to connect with partners, customers and scientists working in a variety of areas, solving difficult scientific problems. Having a full complement of BioTek instruments available at both the BioTek and partner booths allowed for demonstration of possible solutions.

By: BioTek Instruments, Peter J. Brescia Jr., MSc, MBA

Thursday, February 16, 2017

Rigor and Reproducibility

 
Rigor and Reproducibility

A recent report found that the published findings from 89% of preclinical studies in the field of cancer research – in particular, those deemed ‘landmark’ studies – were unable to be confirmed by other scientists [1]. The authors concluded that despite competent, well-intentioned researchers, an alarming number of these inconsistencies were due to limitations in the design and execution of the initial studies, including the use of problematic endpoint measures and biased analysis of data.

This report is not unique to one particular research field, but is in fact representative of a growing awareness within the greater scientific community of the critical need for robust and unbiased experimental design [2-5]. The National Institutes of Health (NIH), one of the world’s leading medical research centers and major provider of extramural biomedical research funds, recently implemented new grant application instructions intended to enhance reproducibility through rigor and transparency [6]. Research grant applicants must now address four areas of focus:  scientific premise, scientific rigor, biological variables, and authentication.

The NIH defines scientific rigor as “the strict application of the scientific method to ensure robust and unbiased experimental design, methodology, analysis, interpretation and reporting of results.” This standard has been particularly difficult to achieve for imaging-based studies, which conventionally depend heavily on manual methods for collecting and analyzing data. This time-consuming approach frequently offers limited quantitative results due to small sample size and qualitative endpoints. Additionally, subjective selection criteria and lack of standardization across methods and instrumentation impedes reproducibility and transparency.

Recent advances in automated imaging systems have provided a great leap forward in our ability to conduct robust imaging-based biomedical research. The efficient and consistent acquisition of high quality images by BioTek’s Lionheart FX and Cytation automated imagers, across large numbers of conditions and replicates, allows researchers to meet the high standards set for reproducibility and transparency. Furthermore, the routine application of powerful Gen5 processing and analysis tools across entire image sets enables robust and meaningful quantitative results. 

Access to innovative technology and the prospect of life-improving therapeutics on the horizon make this an exciting time to be a scientist. Successfully translating biomedical research into clinical advances requires rigorous and reproducible approaches.  In our rapidly evolving scientific landscape, the need to demonstrate the relevance and benefit of sound scientific research is as important now as ever before.

  1. Begley, C.G. and L.M. Ellis, Drug development: Raise standards for preclinical cancer research. Nature, 2012. 483(7391): p. 531-3.
  2. Begley, C.G., A.M. Buchan, and U. Dirnagl, Robust research: Institutions must do their part for reproducibility. Nature, 2015. 525(7567): p. 25-7.
  3. Begley, C.G. and J.P. Ioannidis, Reproducibility in science: improving the standard for basic and preclinical research. Circ Res, 2015. 116(1): p. 116-26.
  4. Kretser, A., D. Murphy, and J. Dwyer, Scientific integrity resource guide: Efforts by federal agencies, foundations, nonprofit organizations, professional societies, and academia in the United States. Crit Rev Food Sci Nutr, 2017. 57(1): p. 163-180.
  5. Pusztai, L., C. Hatzis, and F. Andre, Reproducibility of research and preclinical validation: problems and solutions. Nat Rev Clin Oncol, 2013. 10(12): p. 720-4.
  6. Health, N.I.o. https://grants.nih.gov/reproducibility/index.htm. 2017.


By, BioTek Instruments, Joe Clayton, PhD., Principal Scientist