Journal
NATURE METHODS
Volume 14, Issue 7, Pages 713-+Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nmeth.4305
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Funding
- National Natural Science Foundation of China [31327901, 31521062, 3142800018, 31570839]
- Major State Basic Research Program of China [2013CB531200, 2012CB518200]
- National Science and Technology Major Project Program [2016YFA0500400]
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Developments in miniaturized microscopes have enabled visualization of brain activities and structural dynamics in animals engaging in self-determined behaviors. However, it remains a challenge to resolve activity at single dendritic spines in freely behaving animals. Here, we report the design and application of a fast high-resolution, miniaturized two photon microscope (FHIRM-TPM) that accomplishes this goal. With a headpiece weighing 2.15 g and a hollow-core photonic crystal fiber delivering 920-nm femtosecond laser pulses, the FHIRM-TPM is capable of imaging commonly used biosensors (GFP and GCaMP6) at high spatiotemporal resolution (0.64 urn Laterally and 3.35 mu m axially, 40 Hz at 256 x 256 pixels for raster scanning and 10,000 Hz for free-line scanning). We demonstrate the microscope's robustness with hour-long recordings of neuronal activities at the level of spines in mice experiencing vigorous body movements.
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