Unexpected photobleaching of Alexa 488 in a fixed bacterial sample during 2-photon excitation

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Journal Article
Biotechnic and Histochemistry, 2006, 81 (2-3), pp. 105 - 106
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A sample of fixed bacterial cells was examined by immunofluorescence microscopy using an Alexa 488 conjugated secondary antibody for visualization. Excitation using visible light confirmed the expected photostability of this fluorophore; however, when using 2-photon excitation, Alexa 488 was rapidly and substantially photobleached. The unexpected instability of Alexa 488 under certain conditions may have deleterious consequences if not anticipated and accommodated in experimental protocols.
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