Adaptive Optical Two-Photon Microscopy for Surface-Profiled Living Biological Specimens.
Journal
ACS omega
ISSN: 2470-1343
Titre abrégé: ACS Omega
Pays: United States
ID NLM: 101691658
Informations de publication
Date de publication:
12 Jan 2021
12 Jan 2021
Historique:
received:
06
10
2020
accepted:
05
11
2020
entrez:
18
1
2021
pubmed:
19
1
2021
medline:
19
1
2021
Statut:
epublish
Résumé
We developed adaptive optical (AO) two-photon excitation microscopy by introducing a spatial light modulator (SLM) in a commercially available microscopy system. For correcting optical aberrations caused by refractive index (RI) interfaces at a specimen's surface, spatial phase distributions of the incident excitation laser light were calculated using 3D coordination of the RI interface with a 3D ray-tracing method. Based on the calculation, we applied a 2D phase-shift distribution to a SLM and achieved the proper point spread function. AO two-photon microscopy improved the fluorescence image contrast in optical phantom mimicking biological specimens. Furthermore, it enhanced the fluorescence intensity from tubulin-labeling dyes in living multicellular tumor spheroids and allowed successful visualization of dendritic spines in the cortical layer V of living mouse brains in the secondary motor region with a curved surface. The AO approach is useful for observing dynamic physiological activities in deep regions of various living biological specimens with curved surfaces.
Identifiants
pubmed: 33458495
doi: 10.1021/acsomega.0c04888
pmc: PMC7807736
doi:
Types de publication
Journal Article
Langues
eng
Pagination
438-447Informations de copyright
© 2020 The Authors. Published by American Chemical Society.
Déclaration de conflit d'intérêts
The authors declare no competing financial interest.
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