Leica CW-STED Super-Resolution Demo

In the middle of July 2010 (July 8-July 21; schedule below), the Microscope Facility is pleased to host a demo of Leica's new CW-STED for two-color fluorescence. It's the first super-resolution technique that balances high-speed (5-25 fps) with high resolution (70nm). Normal confocal microscopy has only ~250nm resolution, so CW-STED is ~4x better!

If you are interested in testing this new technology (note restrictions on sample) and help us write a shared instrumentation grant:

1. Immediately: to express interest (if already sent an email, see list at bottom to confirm)
2. Immediately: (see list at bottom to see if your application was received) with 1-3 paragraph description of:
   • How super-resolution would help your research
   • How you’d get appropriate data (for Shared Instrum Grant) from limited 2hr slot
   Examples:
   Morphology: crowded structures (<200nm) not resolvable with std confocal (side-by-side confocal vs CW-STED)
   Dynamics: subcellular motions of small/crowded structures (i.e. not visible in std confocal because of speed or resolution; comparison movies?)
   Colocalization: regulatable multiprotein complexes?  (side-by-side confocal vs CW-STED)
3. Soon: prepare samples for demo:
   Latest details, trade-offs, etc for 2-color sample prep
   Latest details, trade-offs, etc for 1-color sample prep
   

   	Best	Good	AVOID
   Fluor Protein	mCerulean mCitrin mVenus	eCFP eYFP (good for 2-color) eGFP (variable results)
   Dyes	Dylight488 (best for 2-color) Chromeo488	Alexa 488 Atto 488 Oregon Green FITC	DAPI Q-Dots
   2nd color dye	Alexa430 (other color: eYFP OK, but Dylight488 best)
   Mounting Media	Thiodiethanol (see PDFs) + Antifade ProLong (avoid version that contains DAPI) + Antifade	Moviol + Antifade	Slowfade Vectashield

Example of CW-STED

Vimentin-YFP/Keratinocytes (Reiner Windoffer RWTH Aachen Univ.)