Tuesday, April 10, 2018

A New Integrated Metabolic Analysis and Imaging Platform

The theme at this year’s American Association for Cancer Research Annual Meeting (April 14-18) in Chicago is “Driving Innovative Cancer Science to Patient Care”. At the meeting, Agilent Technologies and BioTek Instruments will launch a new integrated solution combining the two companies’ cellular metabolic analysis and imaging technologies.


Agilent’s Seahorse XFe Analyzers simultaneously measure the two major cellular energy-producing pathways, mitochondrial respiration and glycolysis in live cells and in real time. These measurements help scientists better understand the connection between cellular physiological processes and genomic and proteomic data applicable in a variety of applications including cancer research. Cancer cells reprogram their metabolism in order to generate the energy and building blocks they need to proliferate. XF technology provides a window into the Warburg effect, fuel usage and other events that drive tumor cell biology.

For years, Agilent customers have requested an optimized method for verifying assay results and enabling more meaningful data comparisons. BioTek’s Cytation 1 is a cell imaging multi-mode reader offering high contrast brightfield and fluorescence microscopy with up to 60x magnification. When a Cytation is integrated together with an Agilent Seahorse XF Analyzer, the combined platform enables cell biology researchers to fully normalize and analyze samples to answer challenging questions regarding cellular metabolism. Applying a cell count based normalization value makes interpreting XF data and finding relationships amongst the data easier. The integrated solution provides the ability to compare XF data on a well-to-well, plate-to-plate and experiment-to-experiment basis. The ability to incorporate high-quality imagery within Agilent’s Seahorse XF WAVE software adds another dimension to the data. Now researchers can toggle between XF data, brightfield images and fluorescence images in a unified software experience. Referencing images while analyzing XF data provides evidence and guidance on how to limit variability and improve the reproducibility of their XF assays.

The Agilent BioTek collaboration brings to cellular analysis a standardized approach for comparing Seahorse XF data sets and improving assay workflow. For information on the XF Imaging and Normalization System visit www.agilent.com/en/products/cell-analysis/seahorse-xf-imaging-normalization-solution. For information on Cytation 1 Imagers visit www.biotek.com/cytation1.

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

Tuesday, April 3, 2018

Smaller is Better


Sample volume becomes increasingly critical as sample throughput increases for many common applications. Many of these applications, such as protein profiling using LC-MS/MS or RT-PCR, sequencing and micro-array analysis of nucleic acids, rely heavily on accurately determined molecular concentrations of purified biomolecules. Current biomolecular isolation protocols and kits result in yields ranging from nanogram to milligram amounts and are typically eluted in 10-100 μL volumes. Therefore, typical concentrations can range from sub ng/μL to thousands of ng/μL.

Quantification of these purified samples is routinely accomplished by spectrophotometric analysis at 280 nm for proteins and at 260 nm for nucleic acids in a UV transparent vessel. Measurements have traditionally been made in quartz cuvettes with a fixed pathlength of 1 cm and are typically associated with high precision and accurate measurements. Using standard 1 cm pathlength cuvettes, however, often requires diluting nucleic acid samples above about 100-200 ng/μL and protein samples above about 4 mg/mL. This can be less than ideal given the small volume of sample available. Microplates are used to determine analyte concentrations in a more condensed format of 96 samples per plate and lower sample volume. However, a variable pathlength dependent on well diameter and sample volume must be considered during determinations.


More recently, BioTek introduced the use of a micro-volume plate for rapid measurement of multiple undiluted samples with volumes as low as 2 μL, each with 0.5 mm nominal pathlength. The micro-volume plate also provides the capability to make measurements using a cuvette, and/or two vertical 1 cm pathlength vertical cells.

Take3

We have also shown the analytical performance of the plate for micro-volume protein analysis using either native protein absorbance at 280 nm or with the aid of the signal enhancing reagent bicinchoninic acid (BCA). Additionally, we have shown the ability of the accessory to perform micro-volume analysis using UV absorption for both dsDNA and RNA as well the use of the signal enhancing reagent PicoGreen™. Additional methods include monitoring cell growth using turbidity measurements for several cell types including mammalian and yeast cells.

Growth curves of JM109 depicting hourly A600 measurements using either a) 2 µL sample on Take3 accessory plate or b) 500 uL in a low-volume cuvette.

Further information, including a recent webinar highlighting the various methods, can be found at BioTek.com. Under the Applications section, look for Nucleic Acid Quantification or Total Protein Quantification for detailed information. The Resources section contains available on-demand webinars.

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

Friday, March 30, 2018

Call for Awesome Images!


We love to hear about – and see – the wide variety of imaging applications our customers are working with, and we’d love to feature them in BioTek’s 2019 wall calendar. So, we’re having a contest to collect images acquired with BioTek’s Cytation and Lionheart imagers!

Here is your chance to share your best images with your fellow scientists around the world - and the chance to win a cash prize (1st place = $1,000, 2nd place = of $500, 3rd place = $250)!

Click here to enter!



By: BioTek Instruments, Lenore Buehrer, Senior Product Marketing Manager

Tuesday, March 20, 2018

Image integrity and standards in microscopy


When publishing scientific data, raw data is the ground truth and any step taken to manipulate this data should be properly documented. This is especially true with optical microscopy and automated imaging, where software makes it very easy to manipulate digital images. Many online resources document the dos and don’ts of image manipulation for the purpose of scientific publication, for example:

Image integrity and standards
Avoiding Twisted Pixels: Ethical Guidelines for the Appropriate Use and Manipulation of Scientific Digital Images

BioTek Instruments offers a range of automated microscopy and imaging systems with powerful software that can batch-process and analyze thousands of images automatically. Since its early days, BioTek has been involved with human clinical diagnostic applications and because of this, all of our instrumentation and software - even those intended for research - are designed and tested with clinical applications in mind. When we begin designing imaging devices and software, we apply the same principles and make sure that raw images are always preserved with downstream processes documented automatically. This allows tracking any change back to your raw data. Raw data is always available to be re-analyzed as well. As an FDA-registered medical device manufacturer, data integrity is "in our DNA" and you can always trust that microscopic images that you capture on your BioTek devices are traceable, from the raw image all the way to your final cell-level results.

For an overview of what you can do with BioTek's Automated Microscopes and Cell Imaging Systems, please visit our applications page at https://www.biotek.com/applications/imaging-and-microscopy.html.


By: BioTek Instruments, Xavier Amouretti, Manager, Product Marketing

Tuesday, March 13, 2018

Always move forward, but do not forget to look back occasionally

50th Anniversary
This year marks the 50th anniversary for BioTek Instruments. During that time, BioTek has grown from a small garage startup to a leading company in the markets it serves. Much like science, BioTek’s growth was not always a steady progression, but rather a series of rapid leaps, slow growth, and even a few regressions over time. BioTek got its start manufacturing biomedical safety test equipment and later moved into the realm of microplate readers and washer products that served the burgeoning ELISA market in the early ‘80’s. More recently, the company has developed a series of microscopy products. While always moving forward it is important to take a look back at where we have come. Over the last half century, there have been a number of remarkable scientific achievements in which instrumentation designed and manufactured by BioTek played a role. Here is a brief recap of some major discoveries that have shaped the scientific landscape.

Eradication of Smallpox 

In 1980, the World Health Organization announced that smallpox has disappeared worldwide. This infectious disease killed untold millions over the course of centuries, and its eradication through widespread vaccination was a crowning achievement in public health. Smallpox was the first and only infectious disease of humans, to be eradicated by deliberate intervention.

Recombinant DNA Technology 

Recombinant DNA are artificially produced DNA molecules specifically engineered by the combination of genetic material from multiple sources. Recombinant DNA molecules are sometimes called chimeric DNA, because they can be made of material from two different species, like the mythical chimera. In 1978, Werner Arber, Daniel Nathans and Hamilton Smith were awarded the Nobel Prize in Medicine for creating the technology to discover, isolate and apply recombinant DNA. Recombinant genes and proteins are now widely used in all fields of biology.

Polymerase Chain Reaction (PCR) 

Polymerase chain reaction technique for DNA replication was discovered by Kary Mullis, who won a Nobel Prize in 1993 for PCR’s ability to dramatically increase the sensitivity of analysis involving nucleic acids. PCR has become the foundation of modern genetic research, ranging from medicine and evolutionary biology to criminology. PCR-based strategies have propelled huge scientific endeavors such as the Human Genome Project (see below). The technique is widely used by researchers and clinicians to diagnose diseases, clone and sequence genes, and carry out quantitative and genomic studies.

Polymerase Chain Reaction. Schematic representation of logarithmic amplification of DNA sequences with PCR. Curtesy of Wikimedia Commons.

Identification of HIV/AIDS

In 1983 Dr. Françoise Barré-Sinouss and her colleagues at the Pasteur Institute isolated the retrovirus that causes AIDS. The discovery of the human immunodeficiency virus marked the beginning of a continuing effort to develop treatments for a disease that was at the time seen as a death sentence. There is now evidence that it jumped species to humans, probably in the late 19th or early 20th century.

Stem Cells

Stem cells are pluripotent cells that have a capacity to undergo self-renewal and also at the same time have the ability to give rise to at least one or more differentiated or mature cell types. Stem cells are necessary for the production of new and replacement cells for tissues during development and homeostasis, including repair following disease or injury. While scientists originally classified two kinds of stem cells: embryonic stem cells and non-embryonic or adult stem cells, a third type called iPSC has been developed, where differentiated adult cells are reprogramed to be pluripotent. These stem cells can be differentiated into any type of human cell needed for therapeutic purposes.

Decoding of Human Genome

The Human Genome Project started in October of 1990 and was initially headed by Ari Patrinos and Frances Collins. In 2000, scientists from across the world finished a rough draft of the map of the human genome and the final version was realized in 2003, taking more than 10 years and costing about 2.7 billion dollars. The mapped genome shows the placement of every gene and every chromosome that contains all of our genetic material. With the information from individual genome maps, scientists can discover genetic diseases easier. Today, sequencing a human genome costs less than $1000.

Cloning

In 1996 researchers, announce the birth of Dolly, a Finn-Dorset ewe and also the first mammal to be cloned from the adult cell of another animal. She was cloned at the Roslin Institute in Scotland by British scientists Sir Ian Wilmut and Keith Campbell and lived there until her death in 2003 at the age of six. This achievement was followed by a string of other cloned species: ranging from dogs, cats, goats, monkeys and even extinct animal species.

Moon Landing 

In 1969, humans made the first landing on the moon. On July 20, Apollo 11's Neil Armstrong became the first person to step onto the lunar surface. The landing was broadcast on live TV to a worldwide audience, who watched Armstrong step onto the lunar surface and describe the event as "one small step for [a] man, one giant leap for mankind." Buzz Aldrin later followed him. While certainly not related to biology, this event was a watershed moment for myself and countless numbers of others to want to become scientists.
Buzz Aldrin and Lunar lander from Apollo 11 Moon mission. Picture taken by Neil Armstrong. Curtesy of Wikimedia Commons.

My list is certainly not all inclusive, but these advances have stood the test of time in regards to their utility to science. I have only been around for half of BioTek’s journey, but it has been a remarkable journey. I look forward to the future of BioTek and science. To learn about the myriad applications of BioTek Instruments, visit our Applications pages.

By: BioTek Instruments, Paul Held, PhD., Laboratory Manager

Friday, March 2, 2018

Organoids: an endgame for in vitro assay physiological relevance?

My career has been witness to a remarkable progression in the physiological relevance of in vitro assays used in preclinical drug discovery. The 90’s were characterized by industrialized high throughput screening of small chemical compounds for binding to purified proteins. Often millions of chemicals were screened and the protein’s relevance to disease was sketchy, sometimes linked only by their newly discovered DNA sequence.

organoids
10x z-projected image of mouse small intestinal organoid

Times have changed. With the turning of the millennium, these so-called biochemical binding assays were replaced by cell-based assays. At first, easy-to-culture cell lines such as CHO and HEK-293 were used with putative drug target protein overexpressed in them. These have given way to the use of cell types more consistent with the disease (i.e. β-islets for diabetes, neurons and astrocytes for Alzheimer’s). In addition, cells are being grown into structures that resemble tissues using various methods generally described as 3D cell culture.

Yet the most exciting development is the use of organoids, human stem cells that have been coaxed into clusters of differentiated cells that resemble our organs in miniature. As an example, a recent science article provides instructions on how to build a brain in vitro. I think the only way to provide more physiological relevance for an in vitro assay is to miniaturize a human being...

By: BioTek Instruments, Peter Banks Ph.D., Scientific Director

Thursday, March 1, 2018

Multi-Detection, Imaging and HTS Live Cell Applications Workshop

Recently, I had the privilege of leading a multi-day microplate reading and cellular imaging workshop in Santiago, Chile at the Pontificia Universidad Católica de Chile. My friend and colleague, Dr. Carlos F. Lagos, Assistant Professor and one of the leaders at the Center for Translational Endocrinology (CETREN) in the School of Medicine’s Department of Endocrinology, sponsored the event. The purpose of the workshop was to introduce attendees to kinetic and endpoint methods for monitoring multiple phenotypic cellular responses using microscopy and PMT-based detection.

Over the three day workshop, the group used BioTek’s Cytation 5 Cell Imaging Multi-Mode Reader, BioSpa Automated Incubator and MultiFlo FX Multi-Mode Dispenser to perform several assay procedures, including: Cell proliferation, apoptosis, and necrosis using a 2D cell model; Cytotoxicity and cell viability using 2D and 3D cell models; and 2D scratch wound healing assays.

To learn about other applications enabled with Cytation 5, visit BioTek’s Imaging & Microscopy Applications page.

The group is busy with hands-on assay preparation.
Left: Discussing image capture and analysis.
Right: Workshop attendees and representatives from GrupoBios and Pontificia Universidad Catolica de Chile.

By: BioTek Instruments, Brad Larson, Principal Scientist