Tuesday, February 1, 2011

photoconvertable GFP variants for SPT

Many different fluorescent labels are used for super-resolution imaging, and photoconvertable GTP variants are probably the most widely used. And again, there are many photoconvertable GFP variants: PATagRFP, PA-GFP, Dendra, EosFP and many others.

What makes a good photoconvertable GFP for single particle tracking (SPT)? Well, all the things that make a good label (bright, stable, fast to mature, small, inert, this convenient and nicely separated adsorbance and emission spectra - the obvious stuff) and one more thing that is not so obvious.

Photoconvertable label should not convert by itself. It should have a stable dark state and conversion should be strictly upon illumination by the photo-converting light. Otherwise creating single molecules for SPT is hard - they just pop into existence by themselves! In than sense EosFP is much better than Dendra.

Sounds obvious, but it wasn't to me...

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