Abstract:
:Although perfectly transparent specimens are imaged faster with light-sheet microscopy, less transparent samples are often imaged with two-photon microscopy leveraging its robustness to scattering; however, at the price of increased acquisition times. Clearing methods that are capable of rendering strongly scattering samples such as brain tissue perfectly transparent specimens are often complex, costly, and time intensive, even though for many applications a slightly lower level of tissue transparency is sufficient and easily achieved with simpler and faster methods. Here, we present a microscope type that has been geared toward the imaging of semicleared tissue by combining multispot two-photon excitation with rolling shutter wide-field detection to image deep and fast inside semicleared mouse brain. We present a theoretical and experimental evaluation of the point spread function and contrast as a function of shutter size. Finally, we demonstrate microscope performance in fixed brain slices by imaging dendritic spines up to 400-μm deep.
journal_name
J Biomed Optjournal_title
Journal of biomedical opticsauthors
Adam MP,Müllenbroich MC,Di Giovanna AP,Alfieri D,Silvestri L,Sacconi L,Pavone FSdoi
10.1117/1.JBO.23.2.020503subject
Has Abstractpub_date
2018-02-01 00:00:00pages
1-4issue
2eissn
1083-3668issn
1560-2281pii
JBO-170671LRjournal_volume
23pub_type
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