神经元回路的高效多位点双光子功能成像。
Efficient multi-site two-photon functional imaging of neuronal circuits.
作者信息
Castanares Michael Lawrence, Gautam Vini, Drury Jack, Bachor Hans, Daria Vincent R
机构信息
The John Curtin School of Medical Research, Australian National University, Canberra, ACT 2601, Australia.
Research School of Physics and Engineering, Australian National University, Canberra, ACT 2601, Australia.
出版信息
Biomed Opt Express. 2016 Nov 29;7(12):5325-5334. doi: 10.1364/BOE.7.005325. eCollection 2016 Dec 1.
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.
使用高速多通道探测器的双光子成像技术是一种在多个位点对细胞膜动力学进行光学记录的很有前景的方法。该技术的一个主要瓶颈是在短曝光时间（约1毫秒）内捕获的光子数量有限。在此，我们实施时间选通以提高全息投影多个焦点产生的双光子荧光产率，同时保持生物安全的入射平均功率。我们在荧光素和培养的海马神经元中观察到，在诱发钙瞬变时，信噪比（SNR）提高了6倍。随着信噪比的提高，我们可以为实现对响应时间约为1毫秒的荧光探针进行多位点光学记录铺平道路。
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