Tuesday, May 5, 2015

Honey Bee Health and the Critical Link to U.S. Agriculture

Most of us are aware of the role honey bees play in the pollination of plants in the United States but what many don’t realize is how critical they are to commercial crop production. While there are several native pollinators in the U.S., the honey bee was introduced to the New World when European settlers arrived. The honey bee proved to be much more prolific and colonies easy to manage as agriculture expanded across the nation. Honey bees have become indispensible for the production of several crops as evident by the complete dependence of the California almond industry on some 1.4 million colonies, representing ~60% of all colonies in the U.S managed for crop production. Commercial production of several other crops including nuts, fruits, berries and vegetables also depend on pollination by honey bees.
Unfortunately, the U.S. has seen a significant decrease in the total number of managed colonies from ~5 million in the 1940s to 2.5 million today! Along the way the loss and replacement of approximately 10 million bee hives came at a replacement cost of ~ $2 billion. The trend has also been seen on a global scale coinciding with an increased demand for pollination services for commercial operations.

While the disappearance of colonies is well documented, the problem remains a mystery designated Colony Collapse Disorder (CCD). Symptoms of the syndrome include the absence of adult bees or dead bee bodies in the presence of a live queen and brood as well as honey in the hive. The decline in the U.S. has been associated with the introduction of several new pathogens and pests in the 1980s and carriers of viral agents such as Varroa and tracheal mites during the 1990s. However, no single scientific cause has been verified to date leading to CCD. Research suggests rather, that a combination of environmental and pathological stresses can lead to CCD.

Current research has defined the four general categories mentioned above as probable causes contributing to CCD: 1) Pathogens, 2) Parasites, 3) Management stressors and 4) Environmental stressors. Field surveys and laboratory research have supported a number of potential causes yet none have held up to rigorous examination leading to a definitive “cause” of CCD.

One such area of research investigates the affect of the gradual speciation shift in favor of the microsporidia pathogen Nosema ceranae over N. apis via a survey of bee hives across the U.S. Data suggest that infection immunosuppresses honey bees and negatively affects nutrient utilization negatively impacting colony health. Pathogen load via a spore count can be indicative of the health of the colony and generally requires sampling several bees from numerous hives; typically ~ 8-10,000 samples are surveyed over the typical active bee season. Currently the survey relies on manual counting methods similar to blood cell counting using a hemocytometer. BioTek has been working with researchers to provide an automated solution to handle examination of the complex samples of macerated bees. The use of the Cytation 5 at higher magnification (20x) in conjunction with a disposable hemocytometer offers a possible automated solution for determination of spore count (figure 1). Keep tuned for further developments as we continue to work towards validation of this methodology.

Figure 1. Image taken at 20x magnification. Macerated bee sample visualized using phase contrast at 20x magnification. Spores outlined in yellow fit the criteria for intensity, size and circularity while those outlined in red are excluded by non-conformity to the defined circularity criteria

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

Monday, April 27, 2015

The three R’s now spell STEM

It used to be that reading, 'riting and 'rithmetic were the fundamentals of a good education. Now the general consensus is that STEM (science, technology, engineering and math) is the key to success. And boy is it everywhere! Not only are our children exposed to programs at school, they can even attend STEM clubs and camps. For example, some of our awesome BioTek engineers volunteered their time at the 2014-2015 FIRST Tech Robotics Challenge held at the University of Vermont on Pi day (3.14.15). Over 300,000 children competed worldwide for scholarships in this STEM-based competition!

CNN host and writer Fareed Zakaria wrote a recent opinion piece titled "Why America's obsession with STEM education is dangerous". Them's fightin' words Mr. Zakaria! Especially since I’ve been thinking a lot about college lately, for my child that is. And as you may guess, I am one of those parents trying to steer their child towards a STEM major. It’s what I know and mom always knows best, right?

Mr. Zakaria's article got me wondering whether it has to be a left-brain/right-brain world and if you’re a problem-solver, couldn't you be artistic as well? I wouldn't choose the same words as Mr. Zakaria but I understand what he’s trying to say, that a focus on STEM doesn’t guarantee success but rather, one needs the ability to think intelligently and creatively. After all, this is what fosters entrepreneurship and innovation.

By: BioTek Instruments, Ellaine Abueg Ph.D., Product Manager, Specialist

Wednesday, April 15, 2015

Cancer: Emperor of all Maladies

I usually do not watch a great deal of TV, but I am a huge Ken Burns fan. Ken Burns has made a number of very well done documentaries in the past that have aired in the US on public television (PBS). The most notably "The Civil War", which came out 25 years ago, and more recently "The Roosevelts", which aired last year. When I saw that he had directed a new documentary about cancer, I knew that I had to watch it.

The film is a three-part, six hour documentary or "biography" on cancer, based on the Pulitzer Prize winning book "Cancer: The Emperor of all Maladies" written by Siddhartha Kukherjee, an Indian-born American physician and oncologist. The film covers cancer starting from its first documented appearances thousands of years ago through the present.

Episode One: "Magic Bullets" deals with the search for a "cure" for cancer since its discovery thousands of years ago. Its focus centers on Sidney Farber's introduction of chemotherapy in the 1940s.

Episode Two: "The Blind Men and the Elephant" begins in the 1970's with the declaration of the War on Cancer by Richard Nixon.  It deals with the lack of initial success in lowering cancer rates and the desperation for treatments despite the enormous amount of money spent on addressing the disease.  During this time frame the field of molecular biology is born and the Human Genome project is completed.  Only after the gains made in the laboratory spread to the clinic do we see advances in the treatment of some cancers.

Episode Three: "Finding the Achilles Heel" looks at the success and failures of targeted therapies. As the complexity of different cancers becomes unraveled, new avenues of therapy attack become available. However, as with many things in science, with the more knowledge, we realize how much more we need to know.

All three episodes are punctuated with stories of real people afflicted with cancer that really made the series personal to me.  All of us have been touched in some way by cancer and the film certainly portrays that point.  Besides the entertainment and information of the well written narrative, this documentary brings about a bit of nostalgia for me. Much of my Ph.D training took place in a time of great advancement regarding cancer research. Many of the giants of the targeted cancer therapies made their discoveries during those most influential years of my training. The interviews of these researchers, some of whom I've met, all of whom I have great respect for and whose papers I have read and studied years ago reminded me why I chose to pursue a Ph. D degree in the first place.

Having seen the film I must to read the book...

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

Tuesday, April 7, 2015

Cytation - the Sky’s the Limit. Literally.

Time is relative... no surprise there, thanks to Albert Einstein. And comparing time - on a scale as expansive as the Milky Way - to my relatively brief life on Earth makes me feel infinitely small. This feeling is magnified when I visually explore the starry skies. The distance and time between planets, stars and galaxies are mind-boggling.

As a kid, I always questioned: "How many stars are up there?"  I would start counting, reaching several hundred before inevitably giving up in defeat. On a recent work trip to Latin America, I once again considered this question as I enjoyed the night view of the Milky Way over the Arenal Volcano in Costa Rica. There appeared to be thousands upon thousands of stars visible in the clear sky. If only I had a way to count them all...

As I sat there on my balcony, a thought hit me: maybe I could count all the stars using BioTek's Gen5 Image Analysis software, and FINALLY put to rest that question bugging me since childhood! After all, the stars looked VERY similar to how fluorescently labeled cell nuclei look when viewed with BioTek's Cytation Imaging Reader.

So I grabbed my trusty camera out of my luggage, propped it on the balcony ledge and snapped a 30 second long exposure... holding my breath in anticipation. Once the shutter clicked shut, I quickly checked my camera LCD and to my delight each star shone brightly, just like a DAPI-stained cell nucleus.

Opening my laptop, I swiftly downloaded the image and imported it into BioTek's Gen5 software. After just a few clicks of my mouse, I hit "Start" and to my surprise in only 300 ms I had the answer that had eluded me for so many years: 14,115 stars overhead... WOW! That would have taken me forever to count manually!

Maybe this hits home for some of you reading this... those of you still using a handheld clicker counter for counting cells under your microscope, click by click, minute by minute, hour by hour. Think about the hours of your life spent by not automating your image analysis. If Gen5 software can accurately count almost 50,000 stars per second, how much time would it save you in the lab by automating your microscopy and cell counting - which is what Gen5 software was optimized to do!? How much time would it save you? A day, a week, a month... a year?!

The next time you peer through your microscope and start clicking one by one through your cell count, I hope you remember Cytation and Gen5 software. When it comes to challenging applications, Cytation loves stepping up - and the sky’s the limit. Literally!

By: BioTek Instruments, Caleb Foster, Product Manager, Development

Thursday, April 2, 2015

Experimenting with Biology in Bean Town

This week the city of Boston welcomed over 14,000 scientists of various disciplines coming together to discuss current trends in the vast field of experimental biology.  Along with exhibiting at the conference, BioTek hosted our inaugural Cytation™ Product Workshop and Users Group Meeting.  The Cytation series of imaging readers was launched two years ago at EB 2013, uniquely combining automated digital widefield microscopy with conventional multi-mode detection.  Since then, several hundreds of instruments have been installed in customer labs around the world. It has won multiple accolades including a 2013 SelectScience Drug Discovery Product of the Year Award, 2013 MipTec Conference New Product Innovation Award and 2014 Silver Edison Award.  A series of talks were presented by Cytation users describing the research and techniques their labs are performing with Cytation and how it has made a difference in their workflows.  The morning session was kicked off by Joseph Haegele of Cornell University with his talk SCIENCE TAPAS: How one plate has so many uses! Afternoon highlights included Neil Emans of Persomics speaking about miniaturized RNAi screening technology and the imaging of printed libraries with Cytation along with Scott Sneddon of Sharp Edge Labs who demonstrated image cytometry with protein trafficking assays.  The meeting proved to be information packed event full of interaction and constructive dialog between both users and BioTek experts.   Attendees also received a demonstration of BioTek latest release, the Cytation™ 5 Cell Imaging Multi-Mode Reader and imaging enhancements in Gen5™ v2.07 Data Analysis Software. 

By: BioTek Instruments, Jason Greene, Sr. Product Marketing Manager

Tuesday, March 31, 2015

Iceland’s Giant Genome Project: Are there really Genes we can live without?

Nearly 20 years ago a controversial proposal was made to sequence the genomes of the nation of Iceland.  The effort, underwritten by Amgen's deCODE Genetics, a biopharmaceutical company based in Reykjavik, Iceland, hoped to provide scientists insight to the human genome that would expand their ability to investigate the genetic bases of human diseases. Originally the project was granted permission by the Parliament of Iceland, but intervening court cases, citing a right to privacy, limited the unfettered access to patient samples. Despite the privacy concern, the vast majority of the population supported the decode project and over half the population donated blood samples.  Members of this small progressive thinking island nation decided that the scientific benefits of the study to society outweighed any potential prejudice associated with the finding of genetic mutations in individuals.

This small island county is pretty much ideal for genetic studies in that the population is more or less genetically uniform and has been more or less isolated since the first settlers arrived centuries ago.  Equally important is the fact that the health care system of the country maintains health records on all of its citizens.  By sequencing the full genomes of 2,500 Icelanders and comparing the results with less extensive genotype data from more than 104,000 other Icelanders teams of researchers identified 20 million genetic variants.  Keep in mind that the population of Iceland is only 325,000, so nearly 1/3 of the population took part in this study; quite an example of societal cooperation. The researchers then cross-checked that information against Iceland's extensive genealogical and healthcare information records to investigate how diseases might exist through generations of a given family.

The work, published as four papers in the 25 March 2015 issue of the journal Nature Genetics has yielded insights into a number of different areas of genetics, including new insights regarding the common human ancestor for the male Y chromosome or the mutation in the ABCA7 gene, which doubles the risk of Alzheimer’s disease in populations dominated by Icelandic or European ancestry.  One of the more interesting findings was the number of gene deletions or “knockouts” that were found.  More than 1000 genetic knockouts were identified and more than 8,000 Icelanders were identified as having lost the function of at least one gene. Genes responsible for our ability to discriminate smells being the most common knockout, while genes expressed in the brain having far fewer.  While these data seem to demonstrate the redundancy of human genetic architecture, the comparison of phenotypic data with the knockouts can potentially provide information regarding alternative biochemical pathways currently unknown.  Of course the lay-press is now asking whether or not these genes are necessary.

These data have application beyond Iceland.  For example, scientists at deCODE were able to zero in on their new anti-heart-attack compound. It's based on a gene known as LTA4H, first seen in mice, which governs the production of an enzyme called leukotriene A4 hydrolase. The enzyme plays a role in inflammation, a key factor in heart disease, and also encourages the buildup of cholesterol on blood-vessel walls.  Icelanders with a particular variant of the LTA4H gene turn out to be 40% more likely than average to have heart attacks.

While there still exists an ethical conundrum regarding the use of genetic information, it is also obvious that to do nothing with the information might be unethical as well.  How to best use this treasure trove of information to improve their health care is up to the citizens of Iceland.

What do you think of the use of individual genetic information to guide personalized medicine?

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

Tuesday, March 24, 2015

Phenotypic and MoA Analysis of Anti-Metastatic Molecules using 3D Tumoroid Invasion Assays, Kinetic Ligand Binding, and Cellular Microscopy Webinar Recap


Metastasis is the main cause of death in cancer patients and one of the most complex biological processes in human diseases. The development of therapies designed to forestall the metastatic activity of tumors has been met with multiple challenges, including the choice of an appropriate cell model. Tumors in vivo exist as a three-dimensional (3D) mass of multiple cell types, including cancer and stromal cells. Therefore, incorporating a 3D spheroid-type cellular structure that includes co-cultured cell types forming a tumoroid, provides a more predictive model than the use of individual cancer cells cultured on the bottom of a well in traditional two-dimensional (2D) format. A second hurdle has been accurate mechanism of action determination. Possessing knowledge of how invasion is interrupted provides a more complete picture before proceeding to clinical testing. A final hindrance which has been necessary to overcome is the proper capture and analysis of kinetic reader-based data and microscopic images during the tumor invasion process. Being able to monitor inhibitor binding, as well as tumor invasion through the matrix in a quantitative way is critical.

In the webinar presented last week, we demonstrated methods to perform live-cell, label-free, phenotypic analyses of 3D tumor invasion. Spheroid Microplates from Corning Life Sciences, coated with an ultra low attachment surface, were incorporated for tumoroid formation, and performance of the invasion process. Tumor invasion tracking and quantification was then performed via digital microscopy and cellular analysis using the Cytation 5 cell imaging multi-mode reader and Gen5 Data Analysis Software.

During the presentation, we also displayed the ability to determine inhibitor drug-target residence time using the Tag-lite® direct, homogeneous ligand binding assay from Cisbio. The compound set was then incorporated into the tumoroid invasion assay to determine their potential inhibitory effect on the invasion process. Finally, we illustrated how analysis of the involvement of matrix metalloproteinases during the invasion process could be assessed while simultaneously monitoring tumoroid invasion, using the EnSens Protease Activity Detection Technology from Enzium.

The combination of these techniques present accurate, yet easy-to-use methods to assess target-based and phenotypic effects of new, potential anti-metastatic drugs. We invite you to download and watch the recording of the webinar.

By: BioTek Instruments, Brad Larson, Principal Scientist