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

doi:10.1371/journal.pone.0147115

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 journal › Article

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 language	English
Article number	0147115
Number of pages	19
Journal	PLOS One
Volume	11
Issue number	1
DOIs	https://doi.org/10.1371/journal.pone.0147115
Publication status	Published - 29 Jan 2016

Keywords

two-photon
microscopy
femtosecond
Ca2+
hippocampal

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Amor, R., McDonald, A., Trägårdh, J., Robb, G., Wilson, L., Abdul Rahman, N. Z., ... McConnell, G. (2016). Widefield two-photon excitation without scanning: live cell microscopy with high time resolution and low photo-bleaching. PLOS One, 11(1), [0147115]. https://doi.org/10.1371/journal.pone.0147115

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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.",

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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 journal › Article

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AU - Amor, Rumelo

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AU - Robb, Gillian

AU - Wilson, Louise

AU - Abdul Rahman, Nor Zaihana

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AU - Amos, William Bradshaw

AU - Bushell, Trevor J.

AU - McConnell, Gail

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