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.
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.
