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J Biomed Opt. 2015;20(12):126005. doi: 10.1117/1.JBO.20.12.126005.

In vivo wide-field multispectral scanning laser ophthalmoscopy-optical coherence tomography mouse retinal imager: longitudinal imaging of ganglion cells, microglia, and Müller glia, and mapping of the mouse retinal and choroidal vasculature.

Author information

1
University of California Davis, Department of Cell Biology and Human Anatomy, UC Davis RISE Eye-Pod Laboratory, 4320 Tupper Hall, Davis, California 95616, Unites States.
2
Simon Fraser University, School of Engineering Science, 8888 University Drive, Burnaby, British Columbia V5A 1S6, Canada.
3
UC Davis Comprehensive Cancer Center, Department of Biochemistry and Molecular Medicine, 4501 X Street, Sacramento, California 95817, Unites States.
4
University of California Davis, Department of Cell Biology and Human Anatomy, UC Davis RISE Eye-Pod Laboratory, 4320 Tupper Hall, Davis, California 95616, Unites StatescUniversity of California Davis, UC Davis Eye Center, Department of Ophthalmology and V.

Abstract

Scanning laser ophthalmoscopy (SLO) and optical coherence tomography (OCT) provide complementary views of the retina, with the former collecting fluorescence data with good lateral but relatively low-axial resolution, and the latter collecting label-free backscattering data with comparable lateral but much higher axial resolution. To take maximal advantage of the information of both modalities in mouse retinal imaging, we have constructed a compact, four-channel, wide-field (∼50  deg) system that simultaneously acquires and automatically coregisters three channels of confocal SLO and Fourier domain OCT data. The scanner control system allows “zoomed” imaging of a region of interest identified in a wide-field image, providing efficient digital sampling and localization of cellular resolution features in longitudinal imaging of individual mice. The SLO is equipped with a “flip-in” spectrometer that enables spectral “fingerprinting” of fluorochromes. Segmentation of retina layers and en face display facilitate spatial comparison of OCT data with SLO fluorescence patterns. We demonstrate that the system can be used to image an individual retinal ganglion cell over many months, to simultaneously image microglia and Müller glia expressing different fluorochromes, to characterize the distinctive spatial distributions and clearance times of circulating fluorochromes with different molecular sizes, and to produce unequivocal images of the heretofore uncharacterized mouse choroidal vasculature.

PMID:
26677070
PMCID:
PMC4681314
DOI:
10.1117/1.JBO.20.12.126005
[Indexed for MEDLINE]
Free PMC Article

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