In Vivo Imaging of the Fine Structure of Rhodamine-Labeled Macaque Retinal Ganglion Cells

Gray, Daniel C.; Wolfe, Raymond N.; Gee, Bernard P.; Scoles, Drew; Geng, Ying; Masella, Benjamin; Dubra, Alfredo; Luque, S.; Williams, David R.; Merigan, William H.

doi:10.1167/iovs.07-0605

Cited by 61 publications

(53 citation statements)

References 37 publications

(32 reference statements)

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“…It utilized an AO imaging system (Geng et al 2012;Gray et al 2006Gray et al , 2008 to obtain highresolution images of fluorescing cells in retina despite the challenges of low fluorescence levels and eye movements. We employed an UV light source that activates the S-cone opsin to measure ganglion cell responses in the presence of the blue imaging laser, which also excites M-cone opsin and rhodopsin.…”

Section: Discussionmentioning

confidence: 99%

Imaging light responses of retinal ganglion cells in the living mouse eye

Yin

Geng

Osakada

et al. 2013

Journal of Neurophysiology

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This study reports development of a novel method for high-resolution in vivo imaging of the function of individual mouse retinal ganglion cells (RGCs) that overcomes many limitations of available methods for recording RGC physiology. The technique combines insertion of a genetically encoded calcium indicator into RGCs with imaging of calcium responses over many days with FACILE (functional adaptive optics cellular imaging in the living eye). FACILE extends the most common method for RGC physiology, in vitro physiology, by allowing repeated imaging of the function of each cell over many sessions and by avoiding damage to the retina during removal from the eye. This makes it possible to track changes in the response of individual cells during morphological development or degeneration. FACILE also overcomes limitations of existing in vivo imaging methods, providing fine spatial and temporal detail, structure-function comparison, and simultaneous analysis of multiple cells.retinal ganglion cells; in vivo adaptive optics imaging; calcium imaging AVAILABLE METHODS FOR EXAMINING the physiology of retinal ganglion cells (RGCs) have marked limitations, which are resolved by the novel in vivo imaging method described here.

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Section: Discussionmentioning

confidence: 99%

Imaging light responses of retinal ganglion cells in the living mouse eye

Yin

Geng

Osakada

et al. 2013

Journal of Neurophysiology

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“…Gray et al showed that individual RGC bodies, axons, and dendrites could be visualized using adaptive optics retinal imaging in living macaque eyes when a fluorescent dye was introduced into RGCs via retrograde transport after injection into the LGN (Figure 6). 21,3 Ganglion cell bodies, axons, and dendrites labeled with eGFP have also been visualized in living rat eyes using FAOSLO. 29 These techniques are too invasive for use in humans, but many of the studies that are particularly relevant for understanding RGC function are perhaps best applied in animal models.…”

Section: Ganglion Cellsmentioning

confidence: 99%

Imaging retinal mosaics in the living eye

Rossi

Chung

Dubra

et al. 2011

Eye

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Adaptive optics imaging of cone photoreceptors has provided unique insight into the structure and function of the human visual system and has become an important tool for both basic scientists and clinicians. Recent advances in adaptive optics retinal imaging instrumentation and methodology have allowed us to expand beyond cone imaging. Multi-wavelength and fluorescence imaging methods with adaptive optics have allowed multiple retinal cell types to be imaged simultaneously. These new methods have recently revealed rod photoreceptors, retinal pigment epithelium (RPE) cells, and the smallest retinal blood vessels. Fluorescence imaging coupled with adaptive optics has been used to examine ganglion cells in living primates. Two-photon imaging combined with adaptive optics can evaluate photoreceptor function non-invasively in the living primate retina.

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“…However, these contrast with the smooth, bead-free processes of ganglion cell dendrites and axons imaged in vivo (e.g., Gray et al, 2008) and of ganglion cells maintained in organotypic culture . Is bead formation an artifact of tissue processing and, if so, can it be prevented?…”

Section: Introductionmentioning

confidence: 82%

Moniliform deformation of retinal ganglion cells by formaldehyde‐based fixatives

Stradleigh

Greenberg

Partida

et al. 2015

J of Comparative Neurology

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Protocols for characterizing cellular phenotypes commonly use chemical fixatives to preserve anatomical features, mechanically stabilize tissue, and stop physiological responses. Formaldehyde, diluted in either phosphate-buffered saline or phosphate buffer, has been widely used in studies of neurons, especially in conjunction with dyes and antibodies. However, previous studies have reported that these fixatives induce the formation of bead-like varicosities in the dendrites and axons of brain and spinal cord neurons. We report here that these formaldehyde formulations can induce bead formation in the dendrites and axons of adult rat and rabbit retinal ganglion cells, and that retinal ganglion cells differ from hippocampal, cortical, cerebellar, and spinal cord neurons in that bead formation is not blocked by glutamate receptor antagonists, a voltage-gated Na + channel toxin, extracellular Ca 2+ ion exclusion, or temperature shifts. Moreover, we describe a modification of formaldehyde-based fixatives that prevents bead formation in retinal ganglion cells visualized by green fluorescent protein expression and by immunohistochemistry.

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In Vivo Imaging of the Fine Structure of Rhodamine-Labeled Macaque Retinal Ganglion Cells

Cited by 61 publications

References 37 publications

Imaging light responses of retinal ganglion cells in the living mouse eye

Imaging light responses of retinal ganglion cells in the living mouse eye

Imaging retinal mosaics in the living eye

Moniliform deformation of retinal ganglion cells by formaldehyde‐based fixatives

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