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PAmCherry and converted mEos3.2), severely implicating multicolor super-resolution imaging. I have created a mEOS3.2 construct with a mitochondrial target sequence. Importantly, the blue-shifted derivatives emit in the spectral range of the red fluorescent proteins (e. g. The dynamics of this reaction are altered by the presence of reducing agents. In the case of Alexa Fluor 647, the most commonly used fluorescent label in super-resolution microscopy, this derivative is created over time in an intramolecular, irreversible photoinduced chemical reaction. A General Mechanism of Photoconversion of Green-to-Red Fluorescent Proteins Based on. Herein we show that upon intense irradiation with a 561 nm laser line, far-red organic dyes photoconvert to blue-shifted emissive species. Journal of the American Chemical Society, 138(2), 558-565. However, irradiation with high laser intensity can induce photo-conversion of some of the most frequently used fluorophores. We first used the 488-nm laser and green fluorescence of mEos-Orai1 to search and identify cells that exhibited a sparse. A far-red emitting dye is often one of the labels of choice. We took advantage of the ability of mEos3.2 fluorescent protein (Zhang et al., 2012), referred to as mEos hereafter, to photoconvert from the green to the red form and performed single-subunit counting by spot photobleaching. In a multicolor approach, adequate spectral separation of the different photoswitchable probes is required. Localization-based super-resolution microscopy has become an indispensable tool in biology to study features smaller than the diffraction limit of light.