BBB seminar: Joerg Bewersdorf
Pushing the limits of optical nanoscopy: Imaging whole cells at sub-20-nm 3D resolution and living cells in multiple colors
Department of Cell Biology and Department of Biomedical Engineering, Yale University, New Haven, CT, USA
Optical nanoscopy (super-resolution) techniques such as stimulated emission depletion (STED) microscopy and single-molecule switching (SMS) microscopy (e.g. FPALM, PALM or STORM) utilize either targeted or stochastic switching of fluorescent molecules to achieve ~25 nm spatial resolution – about 10-fold below the diffraction limit . The last years have seen many improvements that make these technologies suitable for a rapidly expanding range of applications.
I will report about two recent breakthroughs that significantly expand the live-cell and 3D imaging capabilities of these techniques:
- Optimized instrumentation and labeling approaches now allow multicolor live-cell STED microscopy as a general imaging technique  and allow the observation of morphological changes on the ~50-nm scale.
- A new SMS nanoscope using two opposing objectives (4Pi-SMS/iPALM) which takes advantage of adaptive optics and novel data analysis approaches allows whole-cell imaging at sub-20-nm resolution in 3D .
I will present the scientific foundation, technical realization and application of these new techniques.
(Joerg Bewersdorf has financial interest in Hamamatsu and Bruker.)
 T.J. Gould, S.T. Hess, and J. Bewersdorf “Optical nanoscopy: from acquisition to analysis”, Annu. Rev. Biomed. Eng. 2012; 14:231-54.
 F. Bottanelli, E.B. Kromann, E.S. Allgeyer, R.S. Erdmann, S. Wood Baguley, G. Sirinakis, A. Schepartz, D. Baddeley, D.K. Toomre, J.E. Rothman, and J. Bewersdorf “Two-color live-cell nanoscale imaging of intracellular targets”, Nat. Commun. 2016; 7:10778.
 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, and J. Bewersdorf “Ultra-high resolution 3D imaging of whole cells”, Cell 2016; 166:1028-40.
Chairperson: Jaakko Saraste, Department of Biomedicine