Spatial patterning of regulatory proteins in live Caulobacter crescentus using multicolour super-resolution imaging, now in 3D Seminar

This has permitted access to live cell dynamics at a far greater spatial precision (~10-20nm), much more relevant to the physical interactions of biological systems. These techniques have been exploited to look at the spatial patterning of regulatory proteins in live bacteria.


The photophysical processes underlying the technique will be discussed as well as the importance of chemistry in rational fluorophore design. Finally I will talk about extending super-resolution imaging into three dimensions. Axial information is achieved by the use of a double helix point spread function and illustrated with a fully rendered 3D multicolour live cell super-resolution image of a protein superstructure relative to the cellular membrane.

References

1. Steven F. Lee, Matthew D. Lew, Jerod L. Ptacin, Marissa K. Lee, Robert J. Twieg, Lucy Shapiro, and W. E. Moerner, “Three-dimensional super-resolution co-localization of intracellular protein superstructures and the cell surface in live Caulobacter crescentus,” Proc. Nat. Acad. Sci. (USA), (in press).
2. Steven F. Lee, Michael A. Thompson, Monica Schwartz, Lucy Shapiro, and W. E. Moerner, “Super-Resolution Imaging of the Nucleoid-Associated Protein HU in Caulobacter crescentus,” Biophys. J. Lett. 100, L31-L33 (2011).
3. Jerod L. Ptacin, Steven F. Lee, Ethan C. Garner, Esteban Toro, Michael Eckart, Luis R. Comolli, W.E. Moerner, and Lucy Shapiro, “A spindle-like apparatus guides bacterial chromosome segregation,” Nature Cell Biology 12, 791-798 (2010).
4. Matthew D. Lew, Steven F. Lee, Majid Badieirostami, and W. E. Moerner, “Corkscrew point spread function for far-field three-dimensional nanoscale localization of point objects,” Optics Lett. 36, 202-204 (2011)