I want to use super-resolution microscopy for my project, and am looking at different options to visualize my protein. can you help me with that?
Wendy Stevens Answered Aug 6, 2022
Pennsylvania State University College of Medicine
The choice of fluorophores/dyes used for super-resolution microscopy depends on the technique utilized. For example, STORM and PALM utilize sequential activation of photoswitchable fluorophores to build a time-resolved, high-resolution image and thus requires the use of photoswitchable fluorophores or dyes, such as Alexa-Fluor dyes with different wavelength specs which can be conjugated to an antibody or other protein for imaging. STED, which selectively deactivates fluorophores, can and commonly use Alexa-Fluors, but can also utilize fluorescent proteins such as GFP, or CLIP-/SNAP-tags for in vivo imaging. For SIM, which combines use of a projected periodic grid to create interference patterns with deconvolution, both synthetic dyes such as Alexa-Fluor and genetically encoded fluorophores such as GFP can be used.
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