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Gene. 1996;173(1 Spec No):25-31.

Intravital imaging of green fluorescent protein using two-photon laser-scanning microscopy.

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Beckman Institute (139-74), Division of Biology, California Institute of Technology, Pasadena 91125, USA.


Imaging a fluorophore in a living tissue presents several unique problems. The fluorescence from the labeled cell(s) may be weak, the labeled cells may be buried deep within tissue and the presence of a fluorophore may render the cells photo-sensitive. Two-photon laser-scanning microscopy (TPLSM) offers several advantages in meeting these challenges. We show that TPLSM provides greater sensitivity, better resolution and less photo-bleaching, as compared to confocal laser-scanning microscopy. The dramatically reduced photo-bleaching makes it possible to image cells continuously for long periods of time. Therefore, TPLSM allows a safer and higher-resolution means of imaging living cells labeled with a variety of fluorophores, including green fluorescent protein.

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