Articoli Scientifici di Oftalmologia
Argomento : 32
Adaptive-optics SLO imaging combined with widefield OCT and SLO enables precise 3D localization of fluorescent cells in the mouse retina
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Abstract: Adaptive optics scanning laser ophthalmoscopy (AO-SLO) has
recently been used to achieve exquisite subcellular resolution imaging of
the mouse retina. Wavefront sensing-based AO typically restricts the field
of view to a few degrees of visual angle. As a consequence the relationship
between AO-SLO data and larger scale retinal structures and cellular
patterns can be difficult to assess.
The retinal vasculature affords a largescale
3D map on which cells and structures can be located during in vivo
imaging. Phase-variance OCT (pv-OCT) can efficiently image the
vasculature with near-infrared light in a label-free manner, allowing 3D
vascular reconstruction with high precision. We combined widefield pvOCT
and SLO imaging with AO-SLO reflection and fluorescence imaging
to localize two types of fluorescent cells within the retinal layers: GFPexpressing
microglia, the resident macrophages of the retina, and GFPexpressing
cone photoreceptor cells. We describe in detail a reflective
afocal AO-SLO retinal imaging system designed for high resolution retinal
imaging in mice. The optical performance of this instrument is compared to
other state-of-the-art AO-based mouse retinal imaging systems.
The spatial
and temporal resolution of the new AO instrumentation was characterized
with angiography of retinal capillaries, including blood-flow velocity
analysis. Depth-resolved AO-SLO fluorescent images of microglia and cone
photoreceptors are visualized in parallel with 469 nm and 663 nm
reflectance images of the microvasculature and other structures. Additional
applications of the new instrumentation are discussed.
