Jessica I. Wolfing

In vivo fluorescence imaging of primate retinal ganglion cells and retinal pigment epithelial cells

The ability to resolve single cells noninvasively in the living retina has important applications for the study of normal retina, diseased retina, and the efficacy of therapies for retinal disease. We describe a new instrument for high-resolution, in vivo imaging of the mammalian retina that combines the benefits of confocal detection, adaptive optics, multispectral, and fluorescence imaging. The instrument is capable of imaging single ganglion cells and their axons through retrograde transport in ganglion cells of fluorescent dyes injected into the monkey lateral geniculate nucleus (LGN). In addition, we demonstrate a method involving simultaneous imaging in two spectral bands that allows the integration of very weak signals across many frames despite inter-frame movement of the eye. With this method, we are also able to resolve the smallest retinal capillaries in fluorescein angiography and the mosaic of retinal pigment epithelium (RPE) cells with lipofuscin autofluorescence.

Aberrations induced in wavefront-guided laser refractive surgery due to shifts between natural and dilated pupil center locations

PURPOSE: To determine the aberrations induced in wavefront‐guided laser refractive surgery due to shifts in pupil center location from when aberrations are measured preoperatively (over a dilated pupil) to when they are corrected surgically (over a natural pupil). SETTING: Center for Visual Science and Department of Ophthalmology, University of Rochester, Rochester, New York, USA. METHODS: Shifts in pupil center were measured between dilated phenylephrine hydrochloride (Neo‐Synephrine [2.5%]) and nonpharmacological mesopic conditions in 65 myopic eyes treated with wavefront‐guided laser in situ keratomileusis (Technolas 217z, Bausch & Lomb). Each patients preoperative and 6‐month postoperative wave aberrations were measured over the dilated pupil. Aberrations theoretically induced by decentration of a wavefront‐guided ablation were calculated and compared with those measured 6 months postoperatively (6.0 mm pupil). RESULTS: The mean magnitude of pupil center shift was 0.29 mm ± 0.141 (SD) and usually occurred in the inferonasal direction as the pupil dilated. Depending on the magnitude of shift, the fraction of the higher‐order postoperative root‐mean‐square wavefront error that could be due theoretically to pupil center decentrations was highly variable (mean 0.26 ± 0.20 mm). There was little correlation between the calculated and 6‐month postoperative wavefronts, most likely because pupil center decentrations are only 1 of several potential sources of postoperative aberrations. CONCLUSIONS: Measuring aberrations over a Neo‐Synephrine‐dilated pupil and treating them over an undilated pupil typically resulted in a shift of the wavefront‐guided ablation in the superotemporal direction and an induction of higher‐order aberrations. Methods referencing the aberration measurement and treatment with respect to a fixed feature on the eye will reduce the potential for inducing aberrations due to shifts in pupil center.

Dual-Wavelength Focusing and Simultaneous Image Registration for In Vivo High-Resolution Retinal Imaging

We describe dual-wavelength, simultaneous retinal imaging with compensation for eye movements and monochromatic and chromatic aberrations. Using lipofuscin autofluorescence, we can resolve human retinal pigment epithelial cells in vivo.

In Vivo High-Resolution Fluorescence Retinal Imaging with Adaptive Optics

We describe a new instrument combining adaptive optics ophthalmoscopy and fluorescence imaging. The instrument is capable of imaging retrograde labeled ganglion cells, intrinsic fluorescence from retinal pigment epithelial cells, and intravenous fluorescein injections in vivo.