Efficient multi-site two-photon functional imaging of neuronal circuits

Michael Lawrence Castanares; Vini Gautam; Jack Drury; Hans Bachor; Vincent R. Daria

doi:10.1364/BOE.7.005325

Efficient multi-site two-photon functional imaging of neuronal circuits

Michael Lawrence Castanares, Vini Gautam, Jack Drury, Hans Bachor, Vincent R. Daria^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

16 Citations (Scopus)

Abstract

Two-photon imaging using high-speed multi-channel detectors is a promising approach for optical recording of cellular membrane dynamics at multiple sites. A main bottleneck of this technique is the limited number of photons captured within a short exposure time (~1ms). Here, we implement temporal gating to improve the two-photon fluorescence yield from holographically projected multiple foci whilst maintaining a biologically safe incident average power. We observed up to 6x improvement in the signal-to-noise ratio (SNR) in Fluorescein and cultured hippocampal neurons showing evoked calcium transients. With improved SNR, we could pave the way to achieving multi-site optical recording of fluorogenic probes with response times in the order of ~1ms.

Original language	English
Article number	279008
Pages (from-to)	5325-5334
Number of pages	10
Journal	Biomedical Optics Express
Volume	7
Issue number	12
DOIs	https://doi.org/10.1364/BOE.7.005325
Publication status	Published - 1 Dec 2016

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AU - Castanares, Michael Lawrence

AU - Gautam, Vini

AU - Drury, Jack

AU - Bachor, Hans

AU - Daria, Vincent R.

PY - 2016/12/1

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AB - Two-photon imaging using high-speed multi-channel detectors is a promising approach for optical recording of cellular membrane dynamics at multiple sites. A main bottleneck of this technique is the limited number of photons captured within a short exposure time (~1ms). Here, we implement temporal gating to improve the two-photon fluorescence yield from holographically projected multiple foci whilst maintaining a biologically safe incident average power. We observed up to 6x improvement in the signal-to-noise ratio (SNR) in Fluorescein and cultured hippocampal neurons showing evoked calcium transients. With improved SNR, we could pave the way to achieving multi-site optical recording of fluorogenic probes with response times in the order of ~1ms.

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Efficient multi-site two-photon functional imaging of neuronal circuits

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