Widefield Two-Photon Excitation without Scanning: Live Cell Microscopy with High Time Resolution and Low Photo-Bleaching.
basic_science · Level V
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- Record sourced from PubMed, PMID 26824845.
- Also identified by DOI 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 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.
Medical subject headings
- Hippocampus
- Microscopy, Fluorescence, Multiphoton
- Molecular Imaging
- Neurons
- Time-Lapse Imaging