Widefield Two-Photon Excitation without Scanning: Live Cell Microscopy with High Time Resolution and Low Photo-Bleaching | Zendy

Rumelo Amor | Zendy; Alison McDonald | Zendy; Johanna Trägårdh | Zendy; Gillian Robb | Zendy; Louise Wilson | Zendy; Nor Zaihana Abdul Rahman | Zendy; John Dempster | Zendy; W. B. Amos | Zendy; Trevor J. Bushell | Zendy; Gail McConnell | Zendy

Open Access

Widefield Two-Photon Excitation without Scanning: Live Cell Microscopy with High Time Resolution and Low Photo-Bleaching

Author(s) -

Rumelo Amor,

Alison McDonald,

Johanna Trägårdh,

Gillian Robb,

Louise Wilson,

Nor Zaihana Abdul Rahman,

John Dempster,

W. B. Amos,

Trevor J. Bushell,

Gail McConnell

Publication year - 2016

Publication title -

plos one

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.99

H-Index - 332

ISSN - 1932-6203

DOI - 10.1371/journal.pone.0147115

Subject(s) - two photon excitation microscopy , excitation , microscopy , fluorescence , microscope , fluorescence microscope , photon , optics , materials science , framing (construction) , fluorescence lifetime imaging microscopy , temporal resolution , optoelectronics , chemistry , physics , structural engineering , quantum mechanics , engineering

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.

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