2017
DOI: 10.3791/55579
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Multi-layer Cortical Ca<sup>2+</sup> Imaging in Freely Moving Mice with Prism Probes and Miniaturized Fluorescence Microscopy

Abstract: In vivo circuit and cellular level functional imaging is a critical tool for understanding the brain in action. High resolution imaging of mouse cortical neurons with two-photon microscopy has provided unique insights into cortical structure, function and plasticity. However, these studies are limited to head fixed animals, greatly reducing the behavioral complexity available for study. In this paper, we describe a procedure for performing chronic fluorescence microscopy with cellular-resolution across multipl… Show more

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Cited by 21 publications
(30 citation statements)
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“…Imaging frames were captured at 10.1 frames per second with an exposure time of 49.664 ms at a gain of 2.0. LED power ranged from 50 -80% to adjust the upper tail of the histogram to be as close to a pixel value of 1500 to ensure good signal-to-noise ratio (33). The LED power was set at the beginning of the experiment for each mouse and did not change for the remainder of the experiment.…”
Section: Data Acquisitionmentioning
confidence: 99%
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“…Imaging frames were captured at 10.1 frames per second with an exposure time of 49.664 ms at a gain of 2.0. LED power ranged from 50 -80% to adjust the upper tail of the histogram to be as close to a pixel value of 1500 to ensure good signal-to-noise ratio (33). The LED power was set at the beginning of the experiment for each mouse and did not change for the remainder of the experiment.…”
Section: Data Acquisitionmentioning
confidence: 99%
“…For each animal, movies acquired during baseline and during SD and recovery were concatenated into a timeseries. This timeseries was spatially downsampled by a factor of 2 from 1440 x 1080 pixels to 720 x 540 pixels and temporally downsampled by a factor of 5 to 2 Hz to reduce the data footprint (33). Defective pixels were also rectified (33).…”
Section: Data Processing and Analysismentioning
confidence: 99%
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