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

Pengfei Zhang; Azhar Zam; Yifan Jian; Xinlei Wang; Yuanpei Li; Kit S. Lam; Marie E. Burns; Marinko V. Sarunic; Edward N. Pugh Jr.; Robert J. Zawadzki

doi:10.1117/1.JBO.20.12.126005

16 December 2015 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

Pengfei Zhang, Azhar Zam, Yifan Jian, Xinlei Wang, Yuanpei Li, Kit S. Lam, Marie E. Burns, Marinko V. Sarunic, Edward N. Pugh Jr., Robert J. Zawadzki

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Journal of Biomedical Optics, Vol. 20, Issue 12, 126005 (December 2015). https://doi.org/10.1117/1.JBO.20.12.126005

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 enface 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.

CC BY: © The Authors. Published by SPIE under a Creative Commons Attribution 4.0 Unported License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.

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Pengfei Zhang, Azhar Zam, Yifan Jian, Xinlei Wang, Yuanpei Li, Kit S. Lam, Marie E. Burns, Marinko V. Sarunic, Edward N. Pugh Jr., and Robert J. Zawadzki "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," Journal of Biomedical Optics 20(12), 126005 (16 December 2015). https://doi.org/10.1117/1.JBO.20.12.126005

Published: 16 December 2015

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