Exploring the Exploding Field of Light-Sheet Microscopy at MBL
The underlying concept goes back more than a century, but recent strides in light-sheet microscopy have made it an exceptionally useful tool for biologists investigating living processes, such as embryonic development.
This year, MBL convened its second annual Light Sheet Fluorescence Microscopy (LSFM) conference and workshop to introduce researchers and students to the latest innovations and demonstrate uses of the technology.
The event, held in May, gave 30 attendees a hands-on chance to try out 14 different commercial light-sheet systems. Attendees explored the capabilities and limitations of these systems and learned how to make the best use of them. More than 20 different types of light-sheet microscope systems are currently commercially available, as well as 50 or more 91לhome built91ם systems designed to meet a given lab91יs specific requirements, which alludes to the technology91יs rapid growth.
91לIn the last ten or 12 years, this field has really exploded,91ם said Abhishek Kumar, a Chan-Zuckerberg Initiative imaging scientist at MBL and co-organizer of the event along with , an imaging scientist from Princeton University.
This annual light-sheet workshop supports MBL91יs strategic Imaging Innovation Initiative to develop and facilitate the use of imaging tools for biological research.
91לImaging is an important technological pillar of biomedical science, and we are committed to making the latest technologies accessible and equitable,91ם Kumar said.
A Better Workhorse
Light-sheet microscopy is a viable alternative to confocal microscopy, but it has two great advantages. In traditional confocal microscopy, a tightly focused laser beam scans across a biological sample, stimulating the sample91יs molecules to fluoresce at different identifiable wavelengths. This provides a map of the different proteins and other compounds in the sample.
91לConfocal microscopy is very popular. You go into any biology lab, you91יll find this technique,91ם Kumar said.
But that intense laser beam can damage sensitive samples, and the scanning technique, which illuminates just one point at a time, is slow. Instead, the light-sheet method spreads out the laser light into a plane that can image an entire 2D layer of the sample at a time, making the process much faster. And because this sheet of light is much less intense, it doesn91יt damage delicate samples. As a result, the light-sheet microscopy method has become widespread and is routinely used in biological labs.
91לIn all areas of biology, from live cellular imaging all the way up to corals, people are using light-sheet everywhere,91ם Kumar said. Its advantages over other methods are 91לspeed and gentleness.91ם For example, to produce a 3D image of a large sample using the conventional confocal method might take two days, but with a light-sheet system, just tens of minutes, Kumar said.
Test-Driving an Abundance of Systems
Because there are so many variations of the technology and rapid advances in the field, the conference offered researchers a chance to evaluate the many options available and find those most appropriate for their own specific research.
91לOne system doesn91יt work for all kinds of biological samples,91ם Kumar said.
Light-sheet microscopy has enabled scientists to continuously observe developing embryos in marine organisms over extended periods, without harming them. It is also used to study organs that are normally opaque, but can be chemically altered to become transparent, ranging from mouse brains to frog embryos.
, a microscope developer at the University of Texas-Southwestern and a speaker at the conference, said this technology has been a key tool for his research.
91לIn the realm of live-cell imaging, we are trying to image cancer cells in 3D microenvironments and quantify their shapes and signaling. For that we need fast, 3D microscopy techniques that are gentle to the cells and also possess subcellular 3D resolution,91ם Fiolka said. Those requirements can be met by new light-sheet fluorescence microscopy methods developed by his labs and many others, he said.
, a graduate student at University of Wisconsin-Madison who attended the conference, said, 91לAs someone just beginning their light-sheet journey, I learned so much valuable information 91צ No other microscopy modality is capable of the resolution and penetration into brain tissue that is needed to answer our specific scientific questions.91ם
91לI learned the most from one-on-one conversations with experts regarding issues that I have been having with my own projects back in the lab,91ם Cataldo added. 91לNot only did I receive suggestions on how I could be imaging my mouse brains, but I also received suggestions on stitching, processing, and analyzing91ם the resulting data. Overall, she said, 91לThe conference was highly relevant to my work, and allowed me to gain knowledge on light-sheet fluorescence microscopy, which is a new skill in my research lab.91ם
Fiolka concluded that 91לLSFM is a very dynamic discipline of light microscopy, both in technology, applications, and computational post-processing. LSFM has found a strong foothold in the life sciences, and the emergence of more and more commercial options will likely accelerate its widespread application.91ם
Financial support for the LSFM conference and workshop was generously provided by the commercial vendors, BioImaging of North America, Arnold and Mabel Beckman Foundation, and Chan-Zuckerberg Initiative Imaging Scientists Program.