Biologists now can see more clearly inside cells than ever have been able to before. But there are advantages and challenges to each of the new imaging technologies becoming available. So some of the brightest minds in cell imaging spoke Sunday at an afternoon workshop about the future of fluorescence microscopy and how to choose the right imaging technology for the job.
Peter Kner from the University of Georgia lead off with a discussion of what his lab is doing to improve the resolution of three-dimensional fluorescence imaging. Using adaptive optics borrowed from astronomy to reduce depth distortion, the team is combining different super-resolution techniques – including structured illumination microscopy, which projects a grid pattern on a specimen, and sequential activation techniques like STORM (stochastic optical reconstruction microscopy) — to image model organisms, including C. elegans and zebra fish.
Hari Shroff from the National Institute of Biomedical Imaging and Bioengineering then discussed three-dimensional cell imaging using emerging light sheet technologies. While traditional methods “torture” a cell to death, he noted, light sheet fluorescence microscopy protects specimens from damage and allows rotating them to get different angles of view.
Then Luke Lavis from the Janelia Research Campus of Howard Hughes Medical Institute explained some of the chemistry involved in making the better and brighter fluorescent labels needed for next-generation microscopy. Using rhodamine as a basic scaffold, Lavis’ group synthesizes materials for imaging individual molecules in living cells, which they then distribute free of charge to any researchers who request them.
Justin Taraska from the National Heart, Lung and Blood Institute of the NIH wrapped up by discussing how fluorescent microscopy and electron microscopy combined can produce images beyond the reach of either technology alone. Called correlative light electron microscopy, or CLEM, the combination technology permits the whole-cell imaging of fluorescence as well as the molecular-level resolution of EM.