3D and Multicolor Live-cell Super-resolution Microscopy for Cell Biological Research

Joerg Bewersdorf

Department of Cell Biology, School of Medicine, Yale University, New Haven, CT, USA
 
Cell biological research relies heavily on the capabilities of light microscopes to resolve structures or processes of interest at the nanoscale [1]. My research group is developing super-resolution technology which achieve down to about 20 nm resolution in 3D and applies them to cell biological questions. We are, in particular, focused on improving the 3D resolution, speed, live-cell compatibility, throughput and depth penetration of STED and PALM/STORM microscopes [2, 3].
 
In my presentation, I will provide an overview of recent developments in my lab relevant for cell biological research. I will highlight our unpublished development of multicolor 4Pi-SMS imaging at 20 nm 3D resolution and live-cell applications of STED microscopy to investigate nanoscale dynamics of the endoplasmic reticulum [4].
 
[1] D. Baddeley, J. Bewersdorf “Biological Insight from Super-Resolution Microscopy: What We Can Learn from Localization-Based Images”, Annu. Rev. Biochem 2018 http://www.annualreviews.org/doi/abs/10.1146/annurev-biochem-060815-014801
[2] F. Huang, G. Sirinakis, E.S. Allgeyer, L.K. Schroeder, W.C. Duim, E.B. Kromann, T. Phan, F.E. Rivera-Molina, J.R. Myers, I. Irnov, M. Lessard, Y. Zhang, M.A. Handel, C. Jacobs-Wagner, C.P. Lusk, J.E. Rothman, D.K. Toomre, M.J. Booth, J. Bewersdorf “Ultra-high resolution 3D imaging of whole cells”, Cell 2016 http://www.cell.com/cell/fulltext/S0092-8674(16)30745-0
[3] N. Kilian, A. Goryaynov, M.D. Lessard, G. Hooker, D. Toomre, J.E. Rothman, J. Bewersdorf  “Assessing photodamage in live-cell STED microscopy”, Nature Methods 2018 https://www.nature.com/articles/s41592-018-0145-5
[4] L.K. Schroeder, A.E.S. Barentine, H. Merta, S. Schweighofer, Y. Zhang, D. Baddeley, J. Bewersdorf*, S. Bahmanyar*  “Dynamic nano-scale morphology of the ER surveyed by STED microscopy”, J. Cell Biol 2019 http://jcb.rupress.org/content/218/1/83

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