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3-photon excitation enables fluorescence microscopy deep in densely labeled and highly scattering samples. To date, 3-photon excitation has been restricted to scanning a single focus, limiting the speed of volume acquisition. Here, for the first time to our knowledge, we implemented and characterized dual-plane 3-photon microscopy with temporal multiplexing and remote focusing, and performed simultaneous calcium imaging of two planes beyond 600 µm deep in the cortex of a pan-excitatory GCaMP6s transgenic mouse with a per-plane framerate of 7 Hz and an effective 2 MHz laser repetition rate. This method is a straightforward and generalizable modification to single-focus 3PE systems, doubling the rate of volume (column) imaging with off-the-shelf components and minimal technical constraints.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6865092 | PMC |
http://dx.doi.org/10.1364/BOE.10.005585 | DOI Listing |
Biomed Opt Express
December 2024
The Sainsbury Wellcome Centre for Circuits and Behaviour, University College London, 25 Howland Street, W1T 4JG, London, United Kingdom.
Cortical computations arise from patterns of neuronal activity that span across all cortical layers and cell types. Three-photon excitation has extended the depth limit of imaging within the mouse brain to encompass all cortical layers. However, simultaneous three-photon imaging throughout cortical layers has yet to be demonstrated.
View Article and Find Full Text PDFBiomed Opt Express
November 2019
Allen Institute for Brain Science, 615 Westlake Ave N, Seattle, WA 98109, USA.
3-photon excitation enables fluorescence microscopy deep in densely labeled and highly scattering samples. To date, 3-photon excitation has been restricted to scanning a single focus, limiting the speed of volume acquisition. Here, for the first time to our knowledge, we implemented and characterized dual-plane 3-photon microscopy with temporal multiplexing and remote focusing, and performed simultaneous calcium imaging of two planes beyond 600 µm deep in the cortex of a pan-excitatory GCaMP6s transgenic mouse with a per-plane framerate of 7 Hz and an effective 2 MHz laser repetition rate.
View Article and Find Full Text PDFEnter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!