Taking a Close Look at the Basics of Life with the Help of Lionheart FX
For the curious at heart, science represents an endless reservoir of questions to ask. But sometimes in the quest for knowledge, researchers can overlook the most fundamental yet complex inquiries. This is where Dr. Joseph Larkin aims to make his mark.
At the convergence of Boston University’s Biology and Physics Departments and also as part of the University’s Biological Design Center in Boston, Massachusetts, Dr. Larkin’s new laboratory poses the questions, “How – and why – do living things proactively and reactively interact with each other and their environment to perpetuate?”
As if uncovering answers to those overarching questions wasn’t enough of a challenge, Dr. Larkin also had the distinction of starting up his new lab during the early months of the COVID-19 pandemic in 2020. “I didn’t have a frame of reference as this was my first lab start-up, but it was definitely a challenging time.”
Practical issues, like local stay-at-home advisories as well as equipment and supply backorders, stalled progress. The lack of live personal interaction created another roadblock as it hampered the interpersonal connections necessary to develop a cohesive team and made training of inexperienced team members difficult-to-impossible. On top of these stressors, Dr. Larkin was managing a teaching schedule in a hybrid online and in-person environment.
However, like a macro-scale demonstration of how living things interact and adapt to environmental circumstances, Dr. Larkin and his team ultimately persevered. Today, they are settled into the new lab and happily generating and analyzing experimental data.
Using a soil bacterium and biofilms, the team investigates how microbes engage in multi-cellular group communications and behaviors to change the local environment and enable resiliency under sub-optimal physical and chemical conditions. Using this information, they seek to connect the scale of entire colonies to individual cells.
A significant portion of the lab’s assays revolve around time-lapse microscopy to observe the bacteria growing or moving over substrates. BioTek’s Lionheart FX Automated Microscope is critical in this regard as it is the lab’s primary data collection tool. When Dr. Larkin initially heard of the Lionheart FX, he was a bit hesitant to move away from his perception of traditional microscope technology, but was quite impressed by its power and versatility during a demonstration, and even more impressed once it was installed in the lab.
He is especially taken by Lionheart’s capability to size ratio as it is packed with objectives, fluorescence channels, temperature control, autofocus, and much more. “This compact microscope-in-a-box does about eighty percent of what a massive traditional microscope system does, but with much less space and roughly half the price,” Dr. Larkin says. Lionheart FX doesn’t require a dedicated room, advanced training, or even extensive installation efforts. In fact, time-lapse experiments were being performed using the Lionheart FX on the same day that it was installed.
Using Lionheart FX and 6-well microplates, modified and wild-type bacteria are observed as they grow over 24-hours on a substrate under a variety of conditions. By considering space, time, and behavior, the team can understand how the bacteria sense their environment, and then enter different states of gene expression, including that for selective metabolism, flagella motion, or extracellular matrix production, as examples. The culmination of these activities give colonies their morphology and enable them to change their environment or behave in certain ways.
Lionheart is programmed for the lab’s experiments such that it collects a 5x5 grid of images in each microplate well at pre-determined time points. The images are automatically stitched together using Gen5 Software for easy viewing and analysis.
Team members of all skill levels find the Lionheart FX straightforward and intuitive to use. On top of this, the ability to easily harvest high-quality data helps the less experienced students develop lasting confidence as researchers.
Dr. Larkin summarizes that he is excited to have the Lionheart FX in his lab because he knows that within one day of an experiment, he will have amazing data to contemplate and leverage for future steps.
Figure 1. Cells at the edge of a Bacillus subtilis biofilm that have differentiated into two different states of gene expression, represented by magenta and green captured using the Lionheart FX with a 10X objective. Green: YFP expressed from a motility promoter; magenta: mCherry expressed from an extracellular matrix promoter.
Figure 2. Bacillus subtilis cells collectively swarm over a surface to form branching patterns, captured by Lionheart FX with a 4x objective. Image represents 48 fields of view automatically stitched together. Blue: constitutively expressed CFP.
Figure 3. (L-R) Michael Zulch, Josh Jones, Kimberly Bowal, and Joe Larkin examine data captured using BioTek’s Lionheart FX (background) or Synergy H1 (foreground) in the Larkin Lab at Boston University.
To learn more about Larkin Lab, visit their web site.
Thanks to Dr. Joseph Larkin for sharing his BioTek experience.
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