title: Widefield Two-Photon Excitation without Scanning: Live Cell Microscopy with High Time Resolution and Low Photo-Bleaching.
abstract: We demonstrate fluorescence imaging by two-photon excitation without scanning in biological specimens as previously described by Hwang and co-workers, but with an increased field size and with framing rates of up to 100 Hz. During recordings of synaptically-driven Ca(2+) events in primary rat hippocampal neurone cultures loaded with the fluorescent Ca(2+) indicator Fluo-4 AM, we have observed greatly reduced photo-bleaching in comparison with single-photon excitation. This method, which requires no costly additions to the microscope, promises to be useful for work where high time-resolution is required.
authors.affiliation: Centre for Biophotonics, Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow, United Kingdom.
authors.forename: Rumelo
authors.initials: R
authors.lastname: Amor
authors.affiliation: Centre for Biophotonics, Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow, United Kingdom.
authors.forename: Alison
authors.initials: A
authors.lastname: McDonald
authors.affiliation: Centre for Biophotonics, Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow, United Kingdom.
authors.forename: Johanna
authors.initials: J
authors.lastname: Trägårdh
authors.affiliation: Centre for Biophotonics, Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow, United Kingdom.
authors.forename: Gillian
authors.initials: G
authors.lastname: Robb
authors.affiliation: Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow, United Kingdom.
authors.forename: Louise
authors.initials: L
authors.lastname: Wilson
authors.affiliation: Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow, United Kingdom.
authors.forename: Nor Zaihana
authors.initials: NZ
authors.lastname: Abdul Rahman
authors.affiliation: Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow, United Kingdom.
authors.forename: John
authors.initials: J
authors.lastname: Dempster
authors.affiliation: Centre for Biophotonics, Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow, United Kingdom.|MRC Laboratory of Molecular Biology, Cambridge Biomedical Campus, Cambridge, United Kingdom.
authors.forename: William Bradshaw
authors.initials: WB
authors.lastname: Amos
authors.affiliation: Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow, United Kingdom.
authors.forename: Trevor J
authors.initials: TJ
authors.lastname: Bushell
authors.affiliation: Centre for Biophotonics, Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow, United Kingdom.
authors.forename: Gail
authors.initials: G
authors.lastname: McConnell
mesh_terms: D000814:Aniline Compounds; D000818:Animals; D000831:Animals, Newborn; D002118:Calcium / Q000378:metabolism; D005456:Fluorescent Dyes; D006624:Hippocampus / Q000378:metabolism / Q000648:ultrastructure*; D036641:Microscopy, Fluorescence, Multiphoton / Q000295:instrumentation / Q000379:methods*; D057054:Molecular Imaging / Q000295:instrumentation / Q000379:methods*; D009474:Neurons / Q000378:metabolism / Q000648:ultrastructure*; D038761:Photobleaching; D061251:Primary Cell Culture; D051381:Rats; D017207:Rats, Sprague-Dawley; D013569:Synapses / Q000378:metabolism; D059008:Time-Lapse Imaging / Q000295:instrumentation / Q000379:methods*; D014966:Xanthenes
chemical_list: D000814:Aniline Compounds; C409648:Fluo 4; D005456:Fluorescent Dyes; D014966:Xanthenes; D002118:Calcium
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