Super-resolution microscopy through single molecule localization at cryogenic temperature

• Applicant: Professor Dr. Jörg Enderlein
• Subject Area: Statistical Physics, Nonlinear Dynamics, Complex Systems, Soft and Fluid Matter, Biological Physics ; Biophysics
• Term: from 2017 to 2022
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 390890664

Super-resolution microscopy has revolutionized optical microscopy. In particular, methods based on stochastic fluorophore blinking, such as SOFI, or on single-molecule localization, such as PALM or STORM, have become widely popular. We propose to combine SOFI and PALM with cryo-fluorescence microscopy to enable imaging of shock-frozen samples with high conservation of structural detail and to open the door to correlative cryogenic optical and electron microscopy. A major bottleneck towards these goals is the lack of suitable fluorescent markers able to efficiently photo-switch at cryogenic temperatures, in particular photo-transformable fluorescent proteins (PTFPs). The core goal of our project is to design and characterize PTFPs optimized for cryo-SOFI and cryo-PALM, and to modify our existing cryo-fluorescence microscope for localizing single molecules with extreme precision and allowing cryo-CLEM studies. This research will contribute a new boost in the super-resolution field, of major interest to integrated structural and cell biology.

• DFG Programme: Research Grants
• International Connection: France
• Cooperation Partner: Privatdozent Dr. Dominique Bourgeois, Ph.D.