Wednesday, October 3, 2018

A new Superpower in the Quest for Healing Medicines

Our family recently grew in size… we got a ridiculously adorable puppy that has been keeping us quite busy, both day and night. Truth be told, he is already doing great on most of the typical puppy problem areas – learning to do “his business” outside, sleeping through the night, crate training, etc. The one area where he is still displaying 100% of his playful puppy nature is his chewing and biting. All four sets of hands in our house have punctures and wounds from his teeny-tiny, razor sharp puppy teeth. As I stare down at my hands, it makes me wish there were better medicines out there for quickly healing all my scratches and bites.

In fact, there is a large field of research tied to this exact area - healing wounds - and, the inverse, keeping cells from migrating. One of the leaders in funding wound healing research is the US Department of Defense. They have a keen interest in finding medicines and drugs to improve healing wounds, which would aid dramatically in medicine for warfare. On the other side, the field of oncology is interested in drugs that keep cells from migrating. Drugs with that behavior, specifically targeted at cancer cells, could help treat cancer metastasis and growth. With any research field, scientists work hard to keep all variables in an experiment as standardized and repeatable as possible. In the area of wound healing, one of the key challenges has been standardizing the size and shape of scratches through a layer of cells, to mimic the look and feel of an actual wound. Many researchers today use standard polypropylene pipette tips to create scratches. These do the job, however the scratches through cell monolayers are quite variable. This can create a huge challenge in measurements since the scratches are highly variable in size and shape, which can lead to a significant amount of noise in the final data.

This is where BioTek’s new AutoScratch Wound Making accessory shines (in my mind I have epic music playing while it flies in, stage left). AutoScratch has one purpose in life – to make beautiful, repeatable, make-scientists-smile-with-delight, 800 micron wide scratches through a cell monolayer in either a 24 or 96 well Corning Costar microplate. The innovativeness of the design means that everything is automated, thus removing all the variability that comes from a user making their own scratches. And, for those scientists who hate cleaning up after themselves (I am one of them)... even the cleaning routine is automated! You just can’t beat that!

Automated wound creation using AutoScratch
Manual wound creation using a pipette tip

After AutoScratch exits, stage right, all that is left for the researcher to do is to add drugs looking for either inhibition or activation of wound healing. At the completion of the assay, BioTek offers a number of intuitive imaging systems that will automate the image capture of all the scratches, and will measure key metrics such as the wound width, wound confluence, and max acceleration rate of the wound closure. As I type these last few sentences - with my battered, puppy-scarred hands - my hope is that one of you reading this will have a moment of enlightenment: "Hey, I could use the AutoScratch to discover new wound healing medicines for people just like him". Maybe, just maybe, the next time my family adopts a puppy, there will be a new medicine on the market that heals puppy bites - all enabled by the AutoScratch, this blog, and your ingenious scientific spirit. Please - my hands beg you - don’t delay!

For more information on AutoScratch click here.

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

Tuesday, August 28, 2018

A Matter of Time

When evaluating treatment-induced effects on cells, timing is critical. Cells respond to changes in their environment in diverse ways – from sub-second signaling cascades to changes in cell health that accumulate over days to weeks. The ability to characterize events occurring over these markedly different timescales provides unique insight into cellular processes and increased flexibility for drug development studies.

End-point assays generate a single snapshot of these processes – often at an arbitrarily chosen time point – that is incomplete and easy to misinterpret. In contrast, kinetic imaging-based assays provide a detailed profile of cell characteristics over time, enabling unique quantitative analysis and intuitive validation of results.

BioTek Instrument’s versatile automated imaging systems allow researchers to monitor live cells over a full range of time courses, from seconds to weeks. The instruments maintain optimal environmental growth conditions for long-term studies of cell stress and viability, while an image capture rate of up to 20 frames per second, and aligned dual reagent injectors, enable characterization of rapid cell signaling events.

An example of the unique experiments that are possible with BioTek’s Cytation™ or Lionheart™ FX imaging systems comes from a recent study using expressed biosensors from Montana Molecular. Chemically-induced cell stress by thapsigargin, a potent SERCA pump inhibitor, was measured over a 24 hour period using a new cell stress sensor, while effects on Gq-mediated cell signaling where simultaneously monitored with the R-GECO calcium sensor.

Monitoring stress over 24 hour period in HEK293 cells treated with 1 µM thapsigargin.

These results reveal that cell stress induced by thapsigargin is detected at an order of magnitude lower concentration compared to discernible effects on cellular proliferation. Stress levels peak after 6 hours of treatment. After which, cells either recover naturally from this stress within 22 hours (e.g. 0.1 and 0.3 µM thapsigargin) or progress to more advanced states of distress (i.e. mitotic arrest or cell death).

The effect of thapsigargin on Gq-dependent Ca2+ signaling was probed by adding hM1 receptor agonist to treated cells using the reagent injectors. Ca2+ signaling was reduced in a dose-dependent manner at both 6 and 22 hours during thapsigargin treatment, indicating that the effects of ER stress on Gq-mediated Ca2+ signaling remain even after the cell has shut down the stress response.

Monitoring Gq-dependent calcium signaling in HEK293 cells 6 hours after 1 nM thapsigargin treatment

Click on the image to see larger version.

The ability to monitor cell stress and associated effects on cell signaling is important for understanding a broad range of diseases, as well as for the process of developing drugs designed to treat them. The unique combination of kinetic assays that are possible with BioTek automated imaging systems provides researchers with more physiologically relevant insight and expands research possibilities.

Visit our website to learn more about the full range of application solutions available with BioTek Instruments.

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

Tuesday, August 21, 2018

Alive Wires

In her 2015 Ted Talk The Seafloor is Electric, Laurine Burdorf is speaking about bacteria. Specifically, bacteria that feed on electrons, sending current thru a network of ‘wires’ they grow among themselves. Microbial Fuel Cells (MFCs) are an application of this phenomenon and have undergone a surge in research over the past several years, particularly for their potential role as an alternative energy source. MFCs can already power small electronics, and the variety of suitable microbial ‘power’ strains continue to be discovered all around the world, abundantly, but not exclusively, in dirt, sludge, and mud – hence the seafloor. Although research is ongoing, something these environments share in common is that they are oxygen challenged. One theory is that bacteria adapt to oxygen starved environments by eating electrons then putting out ‘feelers’ to eventually reach bacteria closer to an oxygen source so the electrons can be ‘exhaled’ to continue the energy cycle. A recent trend in the research is investigating whether some bacterial strains can actually sustain viability indefinitely simply by inhaling and exhaling electrons between two electrodes

I recently received and imaged bacilli in a soil sample that had been exposed to an unknown challenge, stained for viability, and then mounted in agar on a microscopy slide. Although not in an electric state, the procedure for imaging and quantitating these bacilli can be universally applied to other strains, and the BioTek Lionheart LX Automated Microscope I used is also amenable to live cell imaging and compatible with a variety of standard and custom vessels such as microchannel substrates. Images were acquired with a 20X objective in both the brightfield and fluorescence channels. A GFP filter cube detected the Syto 9 nucleic acid stain (green), and a PI filter cube detected Propidium Iodide staining for viability (red). Instrument control and analysis was done using BioTek Gen5 Image Prime software. Results were as expected by the scientist that sent the sample.

Quantiative Microscopy Bacilli in soil samples

The basic principles of electric bacteria are available for anyone to learn more about, even using your own dirt. For example, if you are looking for something to do with your kids or grandkids on a rainy vacation day try the MudWatt® Microbe Kit. The more ambitious can build a MFC from your own parts DIY Microbial Fuel Cell - Easy. You can read more and watch a cool video using the link Meet the Electric Life Forms that Live on Pure Energy, a source for some of the information in this blog. If you are a using a BioTek imager in the field of microbiology connect with our Imaging & Microscopy Discussion Group on our Customer Resource Center, and if you don’t have a BioTek Imager, ‘dig in’ and try one!

By: BioTek Instruments, Wendy Goodrich, Applications Scientist

Tuesday, August 14, 2018

AMX™ Automated Media Exchange Module: A New Tool to Simplify Gentle Media Exchanges for Unattached Spheroid Cell Models

It has been widely proven that culturing cells in a three-dimensional (3D) format gives rise to increased cell-cell and cell-matrix interactions and also promotes more biosimilar cell morphologies and behaviors compared to cells cultured on flat 2D surfaces. As such, 3D cell models are being used with increasing frequency for long-term experiments to better mimic in vivo chronic dosing of a test molecule, and in higher density microplate formats to increase throughput. Media exchange and re-dosing steps are critically important during these tests to remove spent media and add fresh media or media with a test molecule. Due to simplicity of cell aggregation and replicate reproducibility, one of the most popular 3D cell culture methods incorporated into long-term test procedures involves the creation of unattached spheroids in media at the bottom of a round-bottom microplate coated to prevent cell attachment. However, with models such as these, care must be taken not to evacuate or damage the spheroid within each well during the exchange process. This can create aspirate and dispense steps that are time consuming and stressful, and can still yield lost spheroids, leading to the loss of critical data.

To alleviate this problem, BioTek has developed a novel peristaltic pump-based automated media exchange method. The AMX™ Automated Media Exchange module for the MultiFlo™ FX uses both available peristaltic pumps, one to dispense and one to aspirate media from the plate wells (Figure 1). The instrument is then programmed to perform the exchange process in a controlled manner that is optimized for the cell type and spheroid size used in each experiment.

MultiFlo FX AMX
Figure 1. MultiFlo FX Multi-Mode Dispenser equipped with the AMX Automated Media Exchange module,
showing aspirate (right arrow) and dispense (left arrow) heads.

During the aspiration step, tubes are positioned to the right of the well center and slightly elevated from the bottom. This allows for only a small residual volume to remain in the well, while ensuring that the spheroid itself is undisturbed (Figure 2).

To replace the removed media in 96-well plates, tubes are again positioned to the right of the well center and slightly elevated from the bottom (Figure 3). For 384-well format, tubes are positioned directly over the spheroid due to the smaller diameter of the well.

These combined aspiration and dispense steps create a method to gently replace spent media either in a single aspirate/dispense procedure for spheroid proliferation assays, or in a multi-step procedure for spheroid washing following fluorescent probe addition or as part of an immunofluorescence staining process.

We invite you to learn more about the AMX module, in addition to the qualitative and quantitative experiments performed to validate its use with multiple cell models and plate types by attending the upcoming webinar:

September 19, 2018
For more information and to register, click here.

By: BioTek Instruments, Brad Larson, Principal Scientist

Tuesday, July 17, 2018

Trading In…or Trading Up?

At home, we’ve been considering trading in our well-loved, well-used, well-running, but old (and lacking up to date technology) vehicle for something newer, easier to operate and with as much of the latest technology as possible within our budget. It’s a tough decision, especially since it still works!

But, we’ll soon find out what the trade-in value is, and probably apply the trade-in value toward a new(er) car, but with improved functionality and useful features to meet our needs.

Many BioTek customers are facing a similar decision today…whether to keep using their old, but still working ELx405™ in the laboratory, or to make the important choice to trade it in toward the purchase of BioTek’s latest technology in microplate washing, the 405™ TS. To make this choice easier, we’re offering ELx405 customers nearly 25% off the purchase of a new 405 TS when they trade in their well-loved, well-used, old, but still working (or not!) ELx405!

405 TS Washer
The 405 TS has many of the same features as the beloved ELx405, but has been updated to meet today’s demanding plate washing workflows, with great benefits including an easy-to-use (even with gloves!) large color touchscreen with intuitive onboard software, patented Ultrasonic Advantage™ for automated manifold cleaning, automated 4-buffer switching and so much more.

Maybe it is time to take advantage of a trade-in offer, and trade up to the 405 TS.

Learn more about this exciting trade-in offer here.

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

Tuesday, July 3, 2018

When will the next big one hit?

Most people feel that the ground that they walk on is a constant. The term “Terra Firma” expounds this belief that the ground is ever stable. Depending on where you live, this is quite true or not so much. We now know that the earth’s outer crust is a series of plates that float on a bed of molten magna and in doing so bump and grind against themselves. There are regions where one plate is diving downward under another, call subduction zones and other areas where two plates are moving laterally. The common feature that the regions all have are earthquakes. Periods where Terra Firma shakes and moves with a tremendous release of energy such that buildings fall and the land is rearranged. These tremors have terrified humankind since the beginning of civilization.

With the advancement of science, we have learned the cause of these titanic events and have worked to try to predict and measure these events. The most common tool are seismometers, which measures the movement of the earth’s crust locally. There sensitivity is such that the epicenter location of earthquakes can be determined and their strength reported as a number on the Richter magnitude scale.

Figure 1. Seismograph tracings from Mexico City around the time of winning goal being scored.
Mother Nature is not the only thing that can cause the Earth to shake. The advent of nuclear weapons has resulted in explosive devices that are detected by the same seismographs that would normally detect Earthquakes. Human activity such as fracking and drilling wastewater disposal, where fluids are injected into the ground to assist the collection of subterranean oil and gas deposits, has resulting in the ground shifting in areas that historically have never had tremors. The state of Oklahoma for example now has more earthquakes than California. Most recently, a potentially human caused tremor was recorded in Mexico City. While not a true Earthquake, some suggest that the Hirving Lozano’s 35th-minute goal in Mexico’s upset victory against defending champion Germany in the 2018 World Cup was the cause. No damage has been reported from the artificial earthquake and Mexico fans are expected to continue celebrating through next week when they face the next round of group stage matches. Mexico will play against South Korea on June 23 at 11 a.m. Eastern.

Now if only the US could manage to qualify we might have our own Earthquake.

By: BioTek Instruments, Paul Held PhD, Laboratory Manager

Tuesday, June 26, 2018

Why so Difficult?

How often have you struggled with a microwave oven you were not familiar with? Maybe at a friend’s place, or at work… Isn’t it amazing how simple tasks become difficult when the interface is not intuitive?

Two weeks ago, I was parking downtown to go to a restaurant with some work guests. We walked to the parking meter and I felt like I was taking an IQ test with three people looking over my shoulder as I was trying to figure out how to pay for parking – I ended up paying until 2 AM by accident! Once, while visiting Amsterdam, I came back to my car blocked with a parking boot after struggling with the parking meter for 10 minutes – talk about a frustrating user experience!

Simple tasks should be easy to accomplish. In the life science world, using a microplate reader should be as simple as walking to the device with your microplate, loading it, starting the read process, getting your data, and done, end-off! Unfortunately, multi-mode microplate readers today are so versatile that simplicity can get lost. Each development iteration adds more hardware features, more software functionality, to a point where running bread and butter assays such as total protein quantification, nucleic acid quantification or ELISA requires taking a number of unnecessary, sometimes complicated steps.
Synergy™ LX Multi-Mode Reader

This is why BioTek developed, the just-launched, Synergy™ LX Multi-Mode Reader. Synergy LX is designed for all the typical microplate-based assays, without the complexity often found in multi-mode plate readers. With absorbance, fluorescence and luminescence detection, a large touch screen, simple interface and USB thumb drive, Synergy LX is the “touch and go” solution for simple multi-detection. And best of all, with Synergy LX, simplicity means budget-friendly!

Don’t struggle with an overly complicated microplate reader.

To learn more on Synergy LX, visit:

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