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Video-rate volumetric neuronal imaging using 3D targeted illumination.

ABSTRACT: Fast volumetric microscopy is required to monitor large-scale neural ensembles with high spatio-temporal resolution. Widefield fluorescence microscopy can image large 2D fields of view at high resolution and speed while remaining simple and costeffective. A focal sweep add-on can further extend the capacity of widefield microscopy by enabling extended-depth-of-field (EDOF) imaging, but suffers from an inability to reject out-of-focus fluorescence background. Here, by using a digital micromirror device to target only in-focus sample features, we perform EDOF imaging with greatly enhanced contrast and signal-to-noise ratio, while reducing the light dosage delivered to the sample. Image quality is further improved by the application of a robust deconvolution algorithm. We demonstrate the advantages of our technique for in vivo calcium imaging in the mouse brain.

SUBMITTER: Xiao S

PROVIDER: S-EPMC5962542 | biostudies-literature | 2018 May

REPOSITORIES: biostudies-literature

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Video-rate volumetric neuronal imaging using 3D targeted illumination.

Xiao Sheng S Tseng Hua-An HA Gritton Howard H Han Xue X Mertz Jerome J

Scientific reports 20180521 1

Fast volumetric microscopy is required to monitor large-scale neural ensembles with high spatio-temporal resolution. Widefield fluorescence microscopy can image large 2D fields of view at high resolution and speed while remaining simple and costeffective. A focal sweep add-on can further extend the capacity of widefield microscopy by enabling extended-depth-of-field (EDOF) imaging, but suffers from an inability to reject out-of-focus fluorescence background. Here, by using a digital micromirror ...[more]

PMID: 29784920

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