Widefield two-photon excitation without scanning: live cell microscopy with high time resolution and low photo-bleaching

Rumelo Amor, Alison McDonald, Johanna Trägårdh, Gillian Robb, Louise Wilson, Nor Zaihana Abdul Rahman, John Dempster, William Bradshaw Amos, Trevor J. Bushell, Gail McConnell

Research output: Contribution to journalArticle

8 Citations (Scopus)
121 Downloads (Pure)

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 Ca2+ events in primary rat hippocampal neurone cultures loaded with the fluorescent Ca2+ 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.
Original languageEnglish
Article number0147115
Number of pages19
JournalPLOS One
Volume11
Issue number1
DOIs
Publication statusPublished - 29 Jan 2016

Keywords

  • two-photon
  • microscopy
  • femtosecond
  • Ca2+
  • hippocampal

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