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
Descripción
Sumario: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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