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

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 hippoc...

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Detalles Bibliográficos
Autores principales: Amor, Rumelo, McDonald, Alison, Trägårdh, Johanna, Robb, Gillian, Wilson, Louise, Abdul Rahman, Nor Zaihana, Dempster, John, Amos, William Bradshaw, Bushell, Trevor J., McConnell, Gail
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2016
Materias:
Research Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4732674/
https://www.ncbi.nlm.nih.gov/pubmed/26824845
http://dx.doi.org/10.1371/journal.pone.0147115
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author Amor, Rumelo
McDonald, Alison
Trägårdh, Johanna
Robb, Gillian
Wilson, Louise
Abdul Rahman, Nor Zaihana
Dempster, John
Amos, William Bradshaw
Bushell, Trevor J.
McConnell, Gail
author_facet Amor, Rumelo
McDonald, Alison
Trägårdh, Johanna
Robb, Gillian
Wilson, Louise
Abdul Rahman, Nor Zaihana
Dempster, John
Amos, William Bradshaw
Bushell, Trevor J.
McConnell, Gail
author_sort Amor, Rumelo
collection PubMed
description 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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spelling pubmed-47326742016-02-04 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 PLoS One Research Article 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. Public Library of Science 2016-01-29 /pmc/articles/PMC4732674/ /pubmed/26824845 http://dx.doi.org/10.1371/journal.pone.0147115 Text en © 2016 Amor et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
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
title Widefield Two-Photon Excitation without Scanning: Live Cell Microscopy with High Time Resolution and Low Photo-Bleaching
title_full Widefield Two-Photon Excitation without Scanning: Live Cell Microscopy with High Time Resolution and Low Photo-Bleaching
title_fullStr Widefield Two-Photon Excitation without Scanning: Live Cell Microscopy with High Time Resolution and Low Photo-Bleaching
title_full_unstemmed Widefield Two-Photon Excitation without Scanning: Live Cell Microscopy with High Time Resolution and Low Photo-Bleaching
title_short Widefield Two-Photon Excitation without Scanning: Live Cell Microscopy with High Time Resolution and Low Photo-Bleaching
title_sort widefield two-photon excitation without scanning: live cell microscopy with high time resolution and low photo-bleaching
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4732674/
https://www.ncbi.nlm.nih.gov/pubmed/26824845
http://dx.doi.org/10.1371/journal.pone.0147115
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