In vivo volumetric imaging of calcium and glutamate activity at synapses with high spatiotemporal resolution

Wei Chen, Ryan G. Natan, Yuhan Yang, Shih-Wei Chou, Qinrong Zhang, Ehud Y. Isacoff and Na Ji ()
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Wei Chen: University of California
Ryan G. Natan: University of California
Yuhan Yang: University of California
Shih-Wei Chou: University of California
Qinrong Zhang: University of California
Ehud Y. Isacoff: University of California
Na Ji: University of California

Nature Communications, 2021, vol. 12, issue 1, 1-12

Abstract: Abstract Studying neuronal activity at synapses requires high spatiotemporal resolution. For high spatial resolution in vivo imaging at depth, adaptive optics (AO) is required to correct sample-induced aberrations. To improve temporal resolution, Bessel focus has been combined with two-photon fluorescence microscopy (2PFM) for fast volumetric imaging at subcellular lateral resolution. To achieve both high-spatial and high-temporal resolution at depth, we develop an efficient AO method that corrects the distorted wavefront of Bessel focus at the objective focal plane and recovers diffraction-limited imaging performance. Applying AO Bessel focus scanning 2PFM to volumetric imaging of zebrafish larval and mouse brains down to 500 µm depth, we demonstrate substantial improvements in the sensitivity and resolution of structural and functional measurements of synapses in vivo. This enables volumetric measurements of synaptic calcium and glutamate activity at high accuracy, including the simultaneous recording of glutamate activity of apical and basal dendritic spines in the mouse cortex.

Date: 2021
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DOI: 10.1038/s41467-021-26965-7

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