MIAP 3D Single-molecule Imaging Workshop

Program

Breaking the resolution limit of conventional microscopy opened the way to investigate cellular structures at the nanoscale, from individual proteins to entire organelles. Different approaches have been proposed from structured illumination microscopy (SIM) to stimulated emission depletion (STED) and single molecule localization microscopy approaches (SMLM). SMLM, such as fluorescence photoactivated localization microscopy (PALM) and stochastic optical reconstruction microscopy (STORM), can provide lateral localization precision down to 10 nm. However, 3D multicolor nanoscopy is still a challenge and a lot of effort has been made by the nano-community to develop quantitative and reproducible 3D super-localization methods.

In this context, abbelight developed a new nanoscope allowing precise isotropic 3D localization precision (15x15x15nm). Two different methods are used to obtain axial information: the supercritical angle fluorescence (SAF) and a strong astigmatism-based PSF measurement. These approaches give rise not only to images resolved at the nanoscale level in 3D, but also to the 3D coordinates of single molecules, opening up new avenues for spatial and temporal quantitative analysis.

The focus of this workshop will be:

• Seminar to discover all theoretical aspects of single-molecule nanoscopy
• Bring your own samples and directly test the technology

This workshop is free of charge but space is limited – be sure to register at your earliest convenience.

Please note:

• If you would like to join a hands-on session and bring your own sample there are certain criteria for your sample preparations. In general, your sample needs to be STORM imaging compatible. For details please see a short guide for sample preparation. If you are not sure, please contact the workshop organizers in advance.
• If you require reagents, antibodies or additional help for your sample preparation, please directly contact the Lighthouse Core Facility, Marie Follo (marie.follo@uniklinik-freiburg.de).