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Published July 1, 1996 | public
Journal Article

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

Abstract

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.

Additional Information

© 1996 Elsevier Science B.V. Received by D.C. Youvan: 1 May 1995; Revised-Accepted: 2 August, 23 August 1995; Received at publishers: 25 September 1995. Thanks to Martie Chalfie and Elizabeth O'Neil for plasmids, Jim Adams for technical assistance, and Larry Zipursky for support. P.G. is a postdoctoral fellow of the Helen Hay Whitney Foundation. Thanks to Sheri McKinney for her technical support with the rat preparations. We thank Molecular Dynamics for providing the Sarastro CLSM and computer hardware used in these studies. The balance of the equipment and the imaging research presented here was supported by a Silvio Conte Center award from NIMH, the NIH Neural Prosthesis Program and support from the Beckman Institute at Caltech.

Additional details

Created:
August 22, 2023
Modified:
October 18, 2023