Two-photon microscopy: Imaging in scattering samples and three-dimensionally resolved flash photolysis: Microscopy research and technique

C Soeller, M B Cannell

Research output: Contribution to journalArticle (Academic Journal)peer-review

51 Citations (Scopus)

Abstract

Two-photon molecular excitation microscopy has several advantages over conventional confocal fluorescence microscopy, including the ability to section deeper into scattering samples and to allow spatially resolved flash photolysis. We describe and examine the benefit of incorporating non-descanned fluorescence detection in our microscope system. In a scattering sample where almost no signal could be obtained at a depth of 50 mu m with confocal detection, non-descanned detection resulted in an improvement of signal strength by more than an order of magnitude at depths >40 mu m. The spatio-temporal properties of stationary spot two-photon excited flash photolysis (TPEFP) in drops of test solutions and cardiac myocytes were also examined. At input powers that produce >10% of the maximum rate of DM-nitrophen photolysis, serious photodestruction of the reporter fluorochrome (Fluo-3) at the photolysis spot occurred. At power levels of similar to 4 mW for periods
Original languageEnglish
Pages (from-to)182-195
Number of pages14
JournalMicroscopy Research and Technique
Volume47
Issue number3
Publication statusPublished - 1 Jan 1999

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