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

6 Citations (Scopus)

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.
LanguageEnglish
Article number0147115
Number of pages19
JournalPLOS One
Volume11
Issue number1
DOIs
Publication statusPublished - 29 Jan 2016

Fingerprint

Photobleaching
photobleaching
Photons
Microscopy
microscopy
Microscopic examination
Scanning
calcium
Optical Imaging
microscopes
Neurons
Rats
Microscopes
neurons
Fluorescence
image analysis
fluorescence
cells
Imaging techniques
rats

Keywords

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

Cite this

Amor, Rumelo ; McDonald, Alison ; Trägårdh, Johanna ; Robb, Gillian ; Wilson, Louise ; Abdul Rahman, Nor Zaihana ; Dempster, John ; Amos, William Bradshaw ; Bushell, Trevor J. ; McConnell, Gail. / Widefield two-photon excitation without scanning : live cell microscopy with high time resolution and low photo-bleaching. In: PLOS One. 2016 ; Vol. 11, No. 1.
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Widefield two-photon excitation without scanning : live cell microscopy with high time resolution and low photo-bleaching. / Amor, Rumelo; McDonald, Alison; Trägårdh, Johanna; Robb, Gillian; Wilson, Louise; Abdul Rahman, Nor Zaihana; Dempster, John; Amos, William Bradshaw; Bushell, Trevor J.; McConnell, Gail.

In: PLOS One, Vol. 11, No. 1, 0147115, 29.01.2016.

Research output: Contribution to journalArticle

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