Pushing the Limits of Optical Nanoscopy


Pushing the Limits of Optical Nanoscopy:  Imaging Whole Cells at Sub-20-nm 3D Resolution and Living Cells in Multiple Colors

Speaker: Joerg Bewersdorf, Department of Cell Biology and Department of Biomedical Engineering, Yale University

Abstract

Optical Nanoscopy (super-resolution) techniques such as STED and FPALM/PALM/STORM (single-molecule switching, SMS) microscopy utilize either targeted or stochastic switching of fluorescent molecules to achieve ~25 nm spatial resolution – about 10-fold below the diffraction limit. [1] 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 [2] 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 [3].

I will present the scientific foundation, technical realization and application of these new techniques.

Joerg Bewersdorf has financial interest in Hamamatsu and Bruker.

[1] T.J. Gould, S.T. Hess, and J. Bewersdorf “Optical Nanoscopy: from Acquisition to Analysis”, Annu. Rev. Biomed. Eng. 2012

[2] 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, J. Bewersdorf “Two-color live-cell nanoscale imaging of intracellular targets”, Nature Communications 2016

[3] 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


This seminar is part of a seminar series hosted by SciLifeLab Fellows

 

Date: Nov 7

Time: 11:00-12:00

Venue: Air&Fire auditorium, SciLifeLab Solna

Host: Ilaria Testa