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Related Experiment Video

Updated: Mar 9, 2026

Targeted Labeling of Neurons in a Specific Functional Micro-domain of the Neocortex by Combining Intrinsic Signal and Two-photon Imaging

Targeted Labeling of Neurons in a Specific Functional Micro-domain of the Neocortex by Combining Intrinsic Signal and Two-photon Imaging

Published on: December 12, 2012

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

Michael Lawrence Castanares¹, Vini Gautam¹, Jack Drury¹

¹The John Curtin School of Medical Research, Australian National University, Canberra, ACT 2601, Australia.

Biomedical Optics Express

|December 27, 2016

Summary

Temporal gating enhances two-photon imaging by improving signal-to-noise ratio for multi-site cellular membrane dynamics recording. This advancement enables faster, clearer optical recordings of neuronal activity.

Keywords:

(110.0110) Imaging systems (170.2520) Fluorescence microscopy

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Related Experiment Videos

Last Updated: Mar 9, 2026

Targeted Labeling of Neurons in a Specific Functional Micro-domain of the Neocortex by Combining Intrinsic Signal and Two-photon Imaging

Targeted Labeling of Neurons in a Specific Functional Micro-domain of the Neocortex by Combining Intrinsic Signal and Two-photon Imaging

Published on: December 12, 2012

Author Spotlight: Comparative Imaging of Neural Activity in Awake and Freely Moving States

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Published on: January 19, 2024

Simultaneous Two-photon In Vivo Imaging of Synaptic Inputs and Postsynaptic Targets in the Mouse Retrosplenial Cortex

Simultaneous Two-photon In Vivo Imaging of Synaptic Inputs and Postsynaptic Targets in the Mouse Retrosplenial Cortex

Published on: March 13, 2016

Area of Science:

Neuroscience
Biophotonics
Cellular Imaging

Background:

Two-photon imaging offers optical recording of cellular membrane dynamics at multiple sites.
Limited photon capture within short exposure times (~1ms) is a key challenge for this technique.
Improving signal-to-noise ratio (SNR) is crucial for high-speed, multi-site imaging.

Purpose of the Study:

To implement temporal gating to enhance two-photon fluorescence yield.
To maintain biologically safe incident average power during imaging.
To improve SNR for multi-site optical recording of cellular dynamics.

Main Methods:

Utilized temporal gating in conjunction with holographic projection of multiple foci.
Performed imaging experiments using Fluorescein and cultured hippocampal neurons.
Measured signal-to-noise ratio improvements under the implemented technique.

Main Results:

Achieved up to a 6x improvement in signal-to-noise ratio (SNR).
Demonstrated enhanced imaging in both Fluorescein and cultured hippocampal neurons.
Observed improved detection of evoked calcium transients.

Conclusions:

Temporal gating effectively boosts fluorescence yield in two-photon imaging.
The developed method enhances SNR, overcoming a major bottleneck in high-speed imaging.
This technique facilitates multi-site optical recording of fluorogenic probes with ~1ms response times.