Invitation to a STED-related mini-seminar

3rd of November 2016, Aud. 2, BBB:

1330-1415    Thommie Karlsson, Application Specialist, Leica Microsystems "Explore Life´s True Nature in Super-Resolution and Nanoscopy - Applications for resolution below the diffraction-limit.."

1430-1530    BBB-seminar: Joerg Bewersdorf, Associate Professor, Yale University, School of Medicine, USA. "Pushing the Limits of Optical Nanoscopy: Imaging Whole Cells at Sub-20-nm 3D Resolution and Living Cells in Multiple Colors."

Abstracts:

Thommie Karlsson: Microscopy beyond the diffraction limit has revolutionized the study of subcellular architecture and dynamics at the nanoscale and is on its way to becoming the new gold standard in fluorescence imaging. The SP8 Hyvolution combines optimal confocal imaging with Huygens from SVI enabling sub-diffraction-limited images with resolution down to 140 nm. Moving towards nanoscopy for further increased resolution STED can also be used enabling resolution below 50nm on the same system. The combination of these techniques allow the user to customize the resolutions toward the applications. Further combinations with advanced imaging equipment enables super resolutions and nanoscopy in all dimensions in deep tissue with multiple colors simultaneously.

Joerg Bewersdorf: 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.