Fixation-resistant photoactivatable fluorescent proteins for CLEM.

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Nature Methods. 2015 Mar;12(3):215-8. doi: 10.1038/nmeth.3225

Fixation-resistant photoactivatable fluorescent proteins for CLEM. Hess LabLooger LabHarris Lab

Paez-Segala MG, Sun MG, Shtengel G, Viswanathan S, Baird MA, Macklin JJ, Patel R, Allen JR, Howe ES, Piszczek G, Hess HF, Davidson MW, Wang Y, Looger LL

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Abstract

Fluorescent proteins facilitate a variety of imaging paradigms in live and fixed samples. However, they lose their fluorescence after heavy fixation, hindering applications such as correlative light and electron microscopy (CLEM). Here we report engineered variants of the photoconvertible Eos fluorescent protein that fluoresce and photoconvert normally in heavily fixed (0.5-1% OsO4), plastic resin-embedded samples, enabling correlative super-resolution fluorescence imaging and high-quality electron microscopy.