Thursday, October 24, 2019

Monarch butterfly: A Beautiful Creature That Feasts on Poison

A recent Facebook post by a work colleague regarding his son being a budding scientist caught my attention. The two of them had found a Monarch butterfly wing in their yard and were looking at it under a microscope.
Monarch
Figure 1.  Female Monarch butterfly.
The Monarch butterfly (Danaus plexippus) is certainly one of the most beautiful of all butterflies and as its name suggests, is considered the king of the butterflies by many. As with all insects, Monarch butterflies go through four stages during one life cycle. The four stages of the Monarch butterfly life cycle are the egg, the larvae (caterpillar), the pupa (chrysalis), and the adult butterfly.
Life cycle stages of the Monarch butterfly
Figure 2.  Life cycle stages of the Monarch butterfly. (Composite of images provided by Wikipedia.)
At the start, the eggs are laid on milkweed plants. They hatch into baby caterpillars, also called the larvae. It takes about four days for the eggs to hatch. Then the baby caterpillar doesn’t do much more than eat the milkweed in order to grow. After about two weeks, the caterpillar will be fully-grown and find a place to attach itself so that it can start the process of metamorphosis. It will attach itself to a stem or a leaf using silk and transform into a chrysalis. Over the 10 days of the chrysalis phase, the body parts of the caterpillar undergo a remarkable transformation, called metamorphosis, to become the beautiful parts of the butterfly. The Monarch butterfly will emerge from the chrysalis and fly away, feeding on flowers for its remaining short life of about two to six weeks.

When I was child, it was fascinating to capture a few Monarch caterpillars from milkweed plants and watch them form a chrysalis and eventually turn into a butterfly. The observations took place at home or in the school classroom.
Figure 3. Milkweed (Asclepias syriaca) showing flowers and latex.
Figure 3. Milkweed (Asclepias syriaca) showing flowers and latex. (Courtesy of Wikipedia)
The interesting point of this is that milkweed (Asclepias), which the larva (caterpillar) feeds on exclusively is quite toxic. Asclepias is a genus of perennial flowering plants known as milkweeds, named for their latex, a milky substance containing cardiac glycosides termed cardenolides, exuded where cells are damaged. Cardiac glycosides affect the sodium (Na+/K+-ATPase) pump of cells. Sodium ion pumps create ion imbalances in cells critical for cardiac and nerve cell function. These are the type of compounds that Agatha Christie featured in a number of her murder mysteries such as Herb of Death, Postern of Fate, and Appointment with Death. Milkweed should kill the caterpillar, but it doesn’t. In fact, the caterpillars store the toxins in their bodies as a defense mechanism against birds that would like to eat them. The bright orange coloring of their wings is actually an “Unsafe to eat” message to animals. So what should kill them actually makes them stronger; the question is how? In a recent article in Nature, researchers used genome editing to retrace the evolution of toxin resistance in the Monarch butterfly. It turns out that only three gene mutations are necessary. These involve amino acid changes at position 111, 119, and 122 of the ATPĪ± subunit of the Na+/K+-ATPase pump. These gene mutations did not occur at once, but rather developed sequentially. Using CRISPR/Cas-9 to edit genes, the group was actually able to make Drosophila fruit flies resistant to milkweed. Studies showed that the mutant flies were 1000 times more resistant to milkweed than the wild type. Using fruit flies, biologists found that these adaptive mutations are not without a cost. It turns out that these mutations had to occur in a specific order. The first mutation, while altering the structure of the pump and conferring some resistance to milkweed also causes neurological problems. The second mutation amended the pump slightly and fixed the neurological problem. This allowed the last mutation, which confers most of the milkweed resistance. The third mutation alone resulted in intolerable neurological seizure issues. Only with the second mutation, would the neurological issues with the third mutation be alleviated.
Figure 4.  The budding scientist at work in his lab.
Figure 4.  The budding scientist at work in his lab.

Oh, back to the budding scientist. His Dad works with me at BioTek, where we manufacture a wide variety of research instrumentation including automated microscopes and imagers, and the software necessary to capture and process image files. He is also a camera buff who is obviously sharing his passion with his son. They managed to take a number of microscopic images of the Monarch butterfly wing, and using BioTek’s Gen5™ software, stitched the individual images into an amazing composite. You can see their work in Figure 5. I think he has a future in science!

Figure 5.   Composite stitched image of wing cells from a Monarch butterfly wing using Gen5.
Figure 5.   Composite stitched image of wing cells from a Monarch butterfly wing using Gen5. 


By: BioTek Instruments, Paul Held PhD, Laboratory Manager

Wednesday, July 31, 2019

Laser Focused and Then Some


When most people hear the word “laser”,  they think of sci-fi thrillers such as Star Wars,  with its hand- held laser blasters, but Albert Einstein proposed the idea of a laser over 100 years ago. Einstein based his theory of stimulated light emission on fundamental physics, more specifically quantum theory. Having recently shown that light was derived of packets of energy (photons), Einstein postulated that if the atoms making up the material are provided with excess energy, individual excited atoms emitting photons could stimulate other excited atoms nearby to do the same. As a result, all photons will have equal energy and move off in the same direction. While the theory was sound, it would take decades before suitable technology was available that would allow the idea to be put into practice. Ultimately, it was shown that when a material is pumped with energy in a mirrored cavity, photons bounce back and forth amplifying the emission of photons. The photons were then allowed to escape through a transparent section in the mirrored surface as a laser beam. Charles Townes at Columbia University produced the first device proving the theory in 1953. The device was capable of amplifying microwaves and was coined the maser.
Tatoute. 2006. Wikipedia.
In 1960, Theodore Maiman, at the Hughes Research Labs in California, produced the first visible-light laser with ruby as the laser medium. However, at this time the laser had few applications,  as did many discoveries stemming from basic research. This was to change toward the  end of the 20th century,  as laser research saw a large expansion and development of high-powered gas, chemical and semiconductor-based lasers. It was not until the development of a laser that worked at room temperature with little or no cooling, that the first widespread use outside of research was realized, in the form of  the compact disk (CD). Today most lasers are of the semiconductor diode type and found throughout industry as well as consumer products of all types. Many lasers have been adapted for biological and biomedical applications ranging from basic research to medical procedures.
Demonstration of a Helium-Neon laser at the Kastler-Brossel Laboratory in Paris. Monniaux, D. 2004. Wikipedia.
Over the past several years, BioTek has implemented lasers in several instruments. The first  was a 680 nm semiconductor laser incorporated into the Synergy Neo for use with PerkinElmer AlphaScreen® technology. BioTek then incorporated a red semiconductor laser as a rapid focusing method during image acquisition for our Cytation™ and Lionheart™ product lines. Most recently, a nitrogen laser operating a 337 nm is an option for the Synergy™ Neo2 Hybrid Multi-Mode Reader
for peak TR-FRET performance. The laser produces approximately 6x more energy than a xenon flash lamp at that wavelength and can flash approximately 2x faster. This results in ideal performance for high throughput screening where both sensitivity and high sample throughput are required. Typical assays include GPCR, kinase, biomarker and cytokine assays using technologies such as Cisbio HTRF®, LanthaScreen™, DELFIA® and LANCE®.
BioTek Instruments, Inc. Synergy Neo2 w/ laser module.



Learn more about BioTek’s patented Laser Autofocus utilized in the Lionheart Automated Microscope and Cytation Cell Imaging Readers.



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

Friday, June 28, 2019

Scratch Assay Starter Kit: Simplifying and Streamlining the Wound Healing Assay

Cell migration is a fundamental biological process important for normal tissue development and pathological processes such as wound healing, tissue repair, and cancer metastasis. Migrating cells undergo coordinated changes in shape and size via carefully orchestrated polymerization and depolymerization of cytoskeletal components. This dramatic rearrangement of cellular components generates motile force and establishes directionality to move the cell along toward its destination. With so many moving parts and biochemical pathways involved, keeping track of the details quickly becomes overwhelming. To model cell migration in vitro, scientists use the well-known scratch assay to measure, probe, and characterize this phenomenon in pursuit of medical research and drug discovery. At first glance the scratch assay is simple; a confluent cell monolayer is physically scraped to leave a gap or wound that the remaining cells can migrate into and “heal”. Look closer though and it is easy to quickly get lost in the details. Whether it is generating uniform and consistent scratches, acquiring high-quality time lapse images, or processing and analyzing images in a robust and objective way, there are many opportunities to introduce variability and prevent accurate quantification.

That’s where BioTek’s Scratch Assay Starter Kit comes in. Consisting of the AutoScratch Wound Making Tool, the Scratch Assay App software, and the necessary cleaning reagents, this kit provides a simple workflow to automate and analyze your scratch assay experiments. At the push of a button, the AutoScratch generates consistent, 800 micron wide scratches in 24-well or 96-well microplates. After scratching, move your plate to a BioTek imager and use the Scratch Assay App to automatically acquire time lapse images of each of your scratches and apply an optimized image analysis. Wound closure rates are tracked and graphed automatically; the healing process is easily visualized with time lapse movies. The Scratch Assay Starter Kit streamlines and automates your assays to increase throughput and ensure reproducibility and accuracy. This lets you focus on the complex biology of cell migration without getting bogged down by data collection and analysis.


Watch BioTek's Webinar on Demand:
A fully automated solution for conducting cell migration assays using the AutoScratch Wound Making Tool
Presenter: Joe Clayton, PhD., Principal Scientist

By: BioTek Instruments, Michael Sfregola, Product Specialist, Imaging

Thursday, April 25, 2019

Black Holes: From the Abstract to Reality

I am a cell biologist at BioTek Instruments, where we manufacture a variety of research instrumentation, some of which are digital microscopes. Like many scientists, I’m also a geek that is interested in fields of science other than my area of research, astronomy for example. One thing about astronomy that has fascinated me is that images of stars in space and cultured cells on a slide look very similar. With astronomy, very large objects (stars) are imaged from very far away. As such, they appear really small and need to be magnified in order to see them. With microscopy, things are very close; the objects are very small and need to be magnified to be seen. Both project as bright objects on a dark background.
Figure 1. NIH3T3 Mouse fibroblasts expressing GFP.
Figure 1. NIH3T3 Mouse fibroblasts expressing GFP.
Then just a few days ago the first images of a black hole was released. Black holes are one of those things you always read about in science fiction novels or see in SciFi movies or shows, not on the front page news. I’m not going to lie; my first thought was it looked like the eye of Sauron in the movie adaptation of The Lord of the Rings.

Image from the Event Horizon Telescope showing the supermassive black hole
Figure 2. Image from the Event Horizon Telescope showing the supermassive black hole in the elliptical galaxy M87, surrounded by superheated material. (EHT Collaboration)
Certainly this is a watershed moment for physics, but it took years of work and the collaboration of hundreds of scientists to make it happen. It also required about half a ton of hard drives. Yes, 1000 pounds of computer hard drives. This is another example of the similarity between microscopy and astronomy - data storage requirements. Before I started working with digital microscopes, my experiments required very little data storage. With digital microscopy, I quickly needed a 1Tb hard drive for my PC, then a 4Tb drive, then a 10Tb drive. Currently, I use a 110 Tb RAID array, but soon that won’t be large enough!

Data collection for the historic black hole image began in 2017 with a coordinated effort called the Event Horizon Telescope (EHT), which is a collection of seven radio telescopes from around the world that are linked to combine the capacity of all those telescopes, creating a “virtual” telescope the size of the Earth.

The now-famous image of a black hole comes from data collected over a period of seven days. At the end of that observation, the EHT didn’t have an image — it had a mountain of data. Scientists at MIT had to develop algorithms to take 5 Petabytes of data and make sense of it. That’s 5000 Terabytes!

While not on the same scale, BioTek Instruments provides Gen5™ as a software tool to combine, process and analyze microscopy images for biomedical research. In the end we hope to provide the tools that our customers need to solve their scientific questions in the microscopic world, but we still salute the cosmic success of MIT and the EHT team.


By: BioTek Instruments, Paul Held PhD, Laboratory Manager

Friday, April 5, 2019

Win Cash, or Better Yet, Bragging Rights! Enter BioTek’s 2019 Imaging Competition


If you've been following us for a while, you probably remember that last year we kicked off our 1st annual Imaging Competition called Imaging Perspectives. Researchers from around the world submitted images captured with their Cytation or Lionheart for a chance to win cash prizes and have their image featured in BioTek's annual wall calendar. Well, the competition proved a success! We loved seeing all of the images and learning about the various applications customers are running with their instruments. So much so that we decided to do it again!

Here’s your chance to show the world what you are working on, and to share the art and beauty that’s often found in science. If you have a favorite image (or three!) that you’ve captured using a BioTek Cytation or Lionheart imager, visit our contest page and submit your entry today! You could win one of three cash prizes (1st place = $1,000, 2nd place = $500, 3rd place = $250) as well as the chance for your image to be featured in our 2020 wall calendar.

To get your creative juices flowing, here are the top three photos from last year:

Imaging Perspectives 2018 Winners

You can click here to see all of our 2018 winners.

Entries for our 2019 competition will be accepted now through the end of July. We can’t wait to see what this year’s submissions will bring!!

Enter now at www.biotek.com/perspectives.

By: BioTek Instruments, Tara Vanderploeg, Marketing Specialist

Friday, February 22, 2019

BioTek Insights User Group Meeting at NIH


On February 7th, BioTek Instruments held its inaugural user’s group meeting, Insights 2019, at the NIH campus in Bethesda, MD. The meeting offered a venue for the exchange of ideas and a chance to develop new collaborations among our customers. Seven BioTek customers presented their research, with topics ranging from novel imaging methods for the screening of drugs inhibiting the sickling of red blood cells, to machine learning for the quantification of morphology changes in zebrafish. Apart from the customer presentations, attendees participated in workshops that demonstrated the following applications:
  • Normalization of Agilent Seahorse OCR and ECAR data using cell counting with Cytation™ 5 
  • Barrier integrity assays using Mimetas OrganoPlate Technology and Cytation 1 
  • Automated media exchange for spheroid proliferation assays using MultiFlo™ FX and the AMX module 
  • Automated scratch assays in 24- and 96-well microplates using AutoScratch™ and Lionheart™ FX
Customers can continue this networking and collaboration through discussion groups on BioTek’s Customer Resource Center (CRC). View discussions and login to post or comment. If you’re not already a CRC user, register with a BioTek instrument serial number or customer number for immediate access.

Many thanks to our presenters and attendees for their participation in this event!

By: BioTek Instruments

Thursday, February 14, 2019

SLAS 2019: Time is money…


The old adage “Time is money” initially came to mind when I walked around the exhibition floor at the 2019 Society for Laboratory Automation and Screening (SLAS) conference in Washington DC. Attendees of this conference have embraced laboratory automation to its full extent. As I perused the floor and talked to different vendors, I couldn’t help but think that all of this laboratory automation equipment was designed for one purpose only: to save time…and time equals money.

It is easy to make this assumption. I used to watch the cartoon The Jetsons growing up and couldn’t help but think how nice it would be if we had Rosie, the robot maid in our house. Imagine all the time we would save by having a robot do our chores! The reason behind laboratory automation is a bit more complex than just saving time.

As a long time field sales representative for BioTek, I have seen my share of researchers make the assumption that laboratory automation is designed solely to help them save time. I would walk into labs and quickly scan the benches to see what was going on. Sometimes I would notice stacks of spent ELISA plates occupying the benches. I would immediately approach the PI or lab manager and asked if they have ever considered a BioTek automated microplate washer or dispenser to help them with their plate washing or dispensing needs. Many of these customers would laugh at me. The conversation would go a bit like this:
Me: I see you are running a lot of ELISA’s in your lab and don’t have a plate washer. How do you wash your plates?

Customer: Oh…we wash them by hand. It is a tedious process that takes a lot of time, but that is the way we have always done this.

Me: Have you ever considered a BioTek automated microplate washer? I see you have a BioTek microplate reader in your lab.

Customer: [Customer chuckles] We have grad students for that and grad student time is cheap. We don’t need a washer.

Me: What if I told you that an automated plate washer will not just save you time but that it can help your lab create more “publishable” results?

Customer: Tell me more….
The main reason behind laboratory automation has less to do with time than it does with consistency and reproducibility. Reproducible results are publishable results. In this example, an automated microplate washer for this customer’s ELISA plates would provide more consistent dispensing and aspiration of wash buffer into the ELISA plate leading to tighter CV’s.

BioTek booth at SLAS 2019

This brings me back to SLAS 2019 and some of the newer technologies we presented there. Systems like the BioTek AutoScratch™ were a big hit at the show. Scratch wound assays can provide cancer researchers with a way of quantifying how different conditions affect cell migration - an important element in the study of cancer metastasis.1 The typical “non-automated” method of creating a scratch in your cell monolayer requires the use of pipette tips and then manually scratching your cells to create a “wound” in the monolayer. Results can be inconsistent when this is done manually - varying downward pressure and scratch inconsistency can result in highly variable results as demonstrated in our recent application note.

After AutoScratch makes the perfect scratch wounds, you can load your assay plates on our BioSpa Live Cell Imaging System or Lionheart™ FX Automated Microscope. BioTek’s Gen5™ software can then use a predefined scratch assay application protocol to image and automatically quantify cell migration because we know exactly where the scratch is on every single well. Our customers at SLAS saw the value in a completely automated workflow solution for this application.

Another hit at SLAS was the new AMX™ Automated Media Exchange module for our MultiFlo™ FX system. Standard plate washers (such as the BioTek 405™ TS or EL406™) are very well established products for washing adherent or lightly adherent cells on microplates. The popularity of 3D cell cultures has required researchers to find new ways of washing non-adherent cells (e.g., spheroids) in microplates. Standard plate washers don’t do well with spheroids as the aspiration pressure would suck the spheroid out of the well. A gentler approach is needed.

I have seen researchers setup wash routines for spheroid washing on complex and expensive pipetting robots. I would describe this as the “killing a fly with a shotgun” approach. You would never use a complex pipetting robot for more standard plate washing routines.

Others have decided to take a step backward and go completely manual with this method; they use hand-held multichannel pipettes to wash their spheroid cultures. I would describe this as the “Karate Kid” approach…or “catching a fly with a set of chopsticks.”

Many of these researchers have explained to me that they have gotten very good with the manual approach and are pretty fast…even faster than the MultiFlo FX with AMX. The problem is consistency and reproducibility. You may miss a well here and there, skip a column on your plate or accidentally aspirate your spheroids when this is done manually. This is where the AMX comes in. It gives you consistent and reproducible results in an automated platform.

We had a great time presenting our new products at SLAS 2019 to our customer base that typically embraces automation. The next time you think about adding automation in your lab, think less about the time savings and more about how automation provides more publishable results!

1. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3857040/


By: BioTek Instruments,  Bikram Chakraborty, Product Manager, Commercial