Document Detail

In vivo non-linear optical (NLO) imaging in live rabbit eyes using the Heidelberg Two-Photon Laser Ophthalmoscope.
MedLine Citation:
PMID:  20558159     Owner:  NLM     Status:  MEDLINE    
Imaging of non-linear optical (NLO) signals generated from the eye using ultrafast pulsed lasers has been limited to the study of ex vivo tissues because of the use of conventional microscopes with slow scan speeds. The purpose of this study was to evaluate the ability of a novel, high scan rate ophthalmoscope to generate NLO signals using an attached femtosecond laser. NLO signals were generated and imaged in live, anesthetized albino rabbits using a newly designed Heidelberg Two-Photon Laser Ophthalmoscope with attached 25 mW fs laser having a central wavelength of 780 nm, pulsewidth of 75 fs, and a repetition rate of 50 MHz. To assess two-photon excited fluorescent (TPEF) signal generation, cultured rabbit corneal fibroblasts (RCF) were first labeled by Blue-green fluorescent FluoSpheres (1 mum diameter) and then cells were micro-injected into the central cornea. Clumps of RCF cells could be detected by both reflectance and TPEF imaging at 6 h after injection. By 6 days, RCF containing fluorescent microspheres confirmed by TPEF showed a more spread morphology and had migrated from the original injection site. Overall, this study demonstrates the potential of using NLO microscopy to sequentially detect TPEF signals from live, intact corneas. We conclude that further refinement of the Two-photon laser Ophthalmoscope should lead to the development of an important, new clinical instrument capable of detecting NLO signals from patient corneas.
Ming Hao; Kevin Flynn; Chyong Nien-Shy; Bryan E Jester; Moritz Winkler; Donald J Brown; Olivier La Schiazza; Josef Bille; James V Jester
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2010-06-15
Journal Detail:
Title:  Experimental eye research     Volume:  91     ISSN:  1096-0007     ISO Abbreviation:  Exp. Eye Res.     Publication Date:  2010 Aug 
Date Detail:
Created Date:  2010-07-20     Completed Date:  2010-08-03     Revised Date:  2014-09-15    
Medline Journal Info:
Nlm Unique ID:  0370707     Medline TA:  Exp Eye Res     Country:  England    
Other Details:
Languages:  eng     Pagination:  308-14     Citation Subset:  IM    
Copyright Information:
Copyright 2010 Elsevier Ltd. All rights reserved.
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MeSH Terms
Cells, Cultured
Cornea / cytology*
Fibroblasts / cytology*
Lasers / diagnostic use*
Microscopy, Fluorescence, Multiphoton / instrumentation,  methods*
Staining and Labeling / methods
Grant Support
EY07348/EY/NEI NIH HHS; EY16663/EY/NEI NIH HHS; R01 EY007348/EY/NEI NIH HHS; R01 EY007348-20S1/EY/NEI NIH HHS; R01 EY007348-22/EY/NEI NIH HHS
Reg. No./Substance:

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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