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TitleDesigning a large field-of-view two-photon microscope using optical invariant analysis.
Publication TypeJournal Article
Year of Publication2018
AuthorsBumstead, Jonathan R., Jasmine J. Park, Isaac A. Rosen, Andrew W. Kraft, Patrick W. Wright, Matthew D. Reisman, Daniel C. Côté, and Joseph P. Culver
Date Published2018 Apr

Conventional two-photon microscopy (TPM) is capable of imaging neural dynamics with subcellular resolution, but it is limited to a field-of-view (FOV) diameter [Formula: see text]. Although there has been recent progress in extending the FOV in TPM, a principled design approach for developing large FOV TPM (LF-TPM) with off-the-shelf components has yet to be established. Therefore, we present a design strategy that depends on analyzing the optical invariant of commercially available objectives, relay lenses, mirror scanners, and emission collection systems in isolation. Components are then selected to maximize the space-bandwidth product of the integrated microscope. In comparison with other LF-TPM systems, our strategy simplifies the sequence of design decisions and is applicable to extending the FOV in any microscope with an optical relay. The microscope we constructed with this design approach can image [Formula: see text] lateral and [Formula: see text] axial resolution over a 7-mm diameter FOV, which is a 100-fold increase in FOV compared with conventional TPM. As a demonstration of the potential that LF-TPM has on understanding the microarchitecture of the mouse brain across interhemispheric regions, we performed imaging of both the cerebral vasculature and microglia cell bodies over the mouse cortex.

Alternate JournalNeurophotonics
PubMed ID29487876
PubMed Central IDPMC5818100
Grant ListR01 NS078223 / NS / NINDS NIH HHS / United States
R01 NS099429 / NS / NINDS NIH HHS / United States