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用于快速采样神经活动荧光报告分子的压缩条纹显微镜技术。

Compressive streak microscopy for fast sampling of fluorescent reporters of neural activity.

作者信息

Cai Changjia, Traubert Owen, Tormes-Vaquerano Jovan, Eybposh M Hossein, Turaga Srinivas C, Rodriguez-Romaguera Jose, Naumann Eva A, Pégard Nicolas C

机构信息

University of North Carolina at Chapel Hill, Department of Applied Physical Sciences, Chapel Hill, North Carolina, United States.

University of North Carolina at Chapel Hill, Joint Department of Biomedical Engineering, Chapel Hill, North Carolina, United States.

出版信息

Neurophotonics. 2025 Apr;12(2):025013. doi: 10.1117/1.NPh.12.2.025013. Epub 2025 May 22.

Abstract

SIGNIFICANCE

one-photon fluorescence imaging of calcium and voltage indicators expressed in neurons enables noninvasive recordings of neural activity with submillisecond precision. However, data acquisition speed is limited by the frame rate of cameras.

AIM

We developed a compressive streak fluorescence microscope to record fluorescence in individual neurons at high speeds ( frames per second) exceeding the nominal frame rate of the camera by trading off spatial pixels for temporal resolution.

APPROACH

Our microscope leverages a digital micromirror device for targeted illumination, a galvo mirror for temporal scanning, and a ridge regression algorithm for fast computational reconstruction of fluorescence traces with high temporal resolution.

RESULTS

In simulations, the ridge regression algorithm reconstructs traces of high temporal resolution with limited signal loss. Validation experiments with fluorescent beads and experiments in larval zebrafish demonstrate accurate reconstruction with a data compression ratio of 10 and accurate recordings of neural activity with 200- to 400-Hz sampling speeds.

CONCLUSIONS

Our compressive microscopy enables new experimental capabilities to monitor activity at a sampling speed that outpaces the nominal frame rate of the camera.

摘要

意义

对神经元中表达的钙和电压指示剂进行单光子荧光成像,能够以亚毫秒级精度对神经活动进行无创记录。然而,数据采集速度受相机帧率限制。

目的

我们开发了一种压缩条纹荧光显微镜,通过用空间像素换取时间分辨率,以超过相机标称帧率的高速(每秒帧数)记录单个神经元中的荧光。

方法

我们的显微镜利用数字微镜器件进行靶向照明,利用振镜进行时间扫描,并利用岭回归算法对具有高时间分辨率的荧光迹线进行快速计算重建。

结果

在模拟中,岭回归算法能在信号损失有限的情况下重建高时间分辨率的迹线。使用荧光珠的验证实验以及在斑马鱼幼体中的实验表明,数据压缩比为10时能实现准确重建,采样速度为200至400赫兹时能准确记录神经活动。

结论

我们的压缩显微镜能够以超过相机标称帧率的采样速度监测活动,从而实现新的实验能力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c17f/12097808/944aab3ae89d/NPh-012-025013-g001.jpg

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