Abstract
We have developed a compact hollow core fiber (HCF)-based imaging platform capable of simultaneous in vivo confocal reflectance and two-photon imaging through the mouse pupil. We demonstrate the performance of this platform by imaging retinal ganglion cells (RGCs) in which the fluorophores YFP and GCaMP3 are expressed in Thy1-YFP-16 and Thy1-GCaMP3 transgenic mice, respectively. Confocal reflectance images of the mouse retina served as a reference for the simultaneous acquisition of the two-photon signals that clearly showed RGCs with single-cell resolution. The use of an HCF platform makes the system compact with future application in the longitudinal investigation into the structure and function of healthy and diseased RGCs.
Highlights
We have developed a compact hollow core fiber (HCF)-based imaging platform capable of simultaneous in vivo confocal reflectance and two-photon imaging through the mouse pupil
Systems used far are bulky and complex and require sophisticated adaptive optics (AO) to compensate for the wavefront errors introduced by the lens and cornea, achieving a tightly focused spot and increased Two-photon excitation (TPE) signal rate
A non-AO two-photon mouse retinal imaging system was described;[9] a water-immersion contact microscope objective lens was used for the focusing optics and the imaging required surgical fixation of the mouse skull
Summary
We have developed a compact hollow core fiber (HCF)-based imaging platform capable of simultaneous in vivo confocal reflectance and two-photon imaging through the mouse pupil. We demonstrate, for the first time to our knowledge, a system based on a hollow core fiber (HCF) that is capable of simultaneous in vivo confocal reflectance and two-photon imaging of RGCs through the mouse pupil without the use of AO.
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