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

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2016 Jan 30

PMID 26824845

Citations 8

Authors

Rumelo Amor

Alison McDonald

Johanna Tragardh

Gillian Robb

Louise Wilson

Nor Zaihana Abdul Rahman

John Dempster

William Bradshaw Amos

Trevor J Bushell

Gail McConnell

Affiliations

Soon will be listed here.

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 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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