ArticleBiomedical optics express2015
Improved two-photon imaging of living neurons in brain tissue through temporal gating.
Article in Biomedical optics express, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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Who cites it
10 citing papers in PubMed.
- Photobleaching Analysis of Fluorescent Proteins in Two-Photon Microscopy at High Repetition Rates.Optics communications · 2025Article
- Pulse train gating to improve signal generation forNeurophotonics · 2023Article
- SNR enhanced high-speed two-photon microscopy using a pulse picker and time gating detection.Scientific reports · 2023Article
- Pulse train gating to improve signal generation forbioRxiv : the preprint server for biology · 2023Article
- Two photon imaging of calcium responses in murine Purkinje neurons.STAR protocols · 2022Article
- Hybrid Electrical and Optical Neural Interfaces.Journal of micromechanics and microengineering : structures, devices, and systems · 2021Article
- Advances in adaptive optics-based two-photon fluorescence microscopy for brain imaging.Lasers in medical science · 2020Review
- Fiber-based tunable repetition rate source for deep tissue two-photon fluorescence microscopy.Biomedical optics express · 2018Article
- Improving Focal Photostimulation of Cortical Neurons with Pre-derived Wavefront Correction.Frontiers in cellular neuroscience · 2017Article
- Efficient multi-site two-photon functional imaging of neuronal circuits.Biomedical optics express · 2016Article
Corrections and comments
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
We optimize two-photon imaging of living neurons in brain tissue by temporally gating an incident laser to reduce the photon flux while optimizing the maximum fluorescence signal from the acquired images. Temporal gating produces a bunch of ~10 femtosecond pulses and the fluorescence signal is improved by increasing the bunch-pulse energy. Gating is achieved using an acousto-optic modulator with a variable gating frequency determined as integral multiples of the imaging sampling frequency. We hypothesize that reducing the photon flux minimizes the photo-damage to the cells. Our results, however, show that despite producing a high fluorescence signal, cell viability is compromised when the gating and sampling frequencies are equal (or effectively one bunch-pulse per pixel). We found an optimum gating frequency range that maintains the viability of the cells while preserving a pre-set fluorescence signal of the acquired two-photon images. The neurons are imaged while under whole-cell patch, and the cell viability is monitored as a change in the membrane's input resistance.
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