An adaptive-optics scanning laser ophthalmoscope for imaging murine retinal microstructure

Abstract

<i>In vivo</i> retinal imaging is an outstanding tool to observe biological processes unfold in real-time. The ability to image microstructure <i>in vivo</i> can greatly enhance our understanding of function in retinal microanatomy under normal conditions and in disease. Transgenic mice are frequently used for mouse models of retinal diseases. However, commercially available retinal imaging instruments lack the optical resolution and spectral flexibility necessary to visualize detail comprehensively. We developed an adaptive optics scanning laser ophthalmoscope (AO-SLO) specifically for mouse eyes. Our SLO is a sensor-less adaptive optics system (no Shack Hartmann sensor) that employs a stochastic parallel gradient descent algorithm to modulate a deformable mirror, ultimately aiming to correct wavefront aberrations by optimizing confocal image sharpness. The resulting resolution allows detailed observation of retinal microstructure. The AO-SLO can resolve retinal microglia and their moving processes, demonstrating that microglia processes are highly motile, constantly probing their immediate environment. Similarly, retinal ganglion cells are imaged along with their axons and sprouting dendrites. Retinal blood vessels are imaged both using evans blue fluorescence and backscattering contrast.