Longitudinal adaptive optics fluorescence microscopy reveals cellular mosaicism in patients

Jung, Hyunwoo; Liu, Jianfei; Liu, Tao; George, Aman; Smelkinson, Margery; Cohen, Sarah; Sharma, Ruchi; Schwartz, Owen; Maminishkis, Arvydas; Bharti, Kapil; Cukras, Catherine A; Huryn, Laryssa A.; Brooks, Brian P.; Fariss, Robert N; Tam, Johnny

doi:10.1172/jci.insight.124904

Cited by 26 publications

(17 citation statements)

References 50 publications

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“…Whilst immune cell labelling was the central objective, we illustrate a range of retinal pathology can be imaged late after ICG administration, with distinctive signals arising from the RPE. DNIRA has been described before, but here we demonstrate findings across a further diverse range of human ocular pathology using a commercially available SLO platform [9] that complements recent findings using adaptive optics imaging of the RPE in inherited retinal degenerations [8]. Several examples of unique patterns only seen by DNIRA identify additional or complementary information for monitoring RPE health versus traditional 488 nm autofluorescence imaging.…”

Section: Discussionsupporting

confidence: 67%

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Intravenous indocyanine green dye is insufficient for robust immune cell labelling in the human retina

et al. 2020

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It is not currently possible to reliably visualise and track immune cells in the human central nervous system or eye. Previous work demonstrated that indocyanine green (ICG) dye could label immune cells and be imaged after a delay during disease in the mouse retina. We report a pilot study investigating if ICG can similarly label immune cells within the human retina. Twelve adult participants receiving ICG angiography as part of routine standard of care were recruited. Baseline retinal images were obtained prior to ICG administration then repeated over a period ranging from 2 hours to 9 days. Matched peripheral blood samples were obtained to examine systemic immune cell labelling and activation from ICG by flow cytometry with human macrophage cultures as positive controls. Differences between the delayed near infrared ICG imaging and 488 nm autofluorescence was observed across pathologies, likely arising from the retinal pigment epithelium (RPE). Only one subject demonstrated ICG signal on peripheral blood myeloid cells and only three distinct cell-sized signals appeared over time within the retina of three participants. No significant increase in immune cell activation markers were detected after ICG administration. ICG accumulated in the endosomes of macrophage cultures and was detectable above a minimum concentration, suggesting cell labelling is possible. ICG can label RPE and may be used as an additional biomarker for RPE health across a range of retinal disorders. Standard clinical doses of intravenous ICG do not lead to robust immune cell labelling in human blood or retina and further optimisation in dose and route are required.

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Section: Discussionsupporting

confidence: 67%

“…ICG is known to enhance the visibility of the retinal pigment epithelium (RPE) mosaic and has been used in adaptive optics imaging [7,8]. This staining has also been seen late after administration in the normal rat retina [9] in an approach termed delayed near-infra red analysis (DNIRA).…”

Section: Introductionmentioning

confidence: 99%

Intravenous indocyanine green dye is insufficient for robust immune cell labelling in the human retina

et al. 2020

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“…More recently, AO was combined with swept-source OCT to achieve deeper tissue penetration (144). Additionally, fluorescence imaging techniques have been described in humans and animals for enhanced visualisation of retinal cells and microvasculature (151). Two-photon excitation fluorescence has also been widely used in animal models to capture high-contrast images of retinal structure (47).…”

Section: Future Of Adaptive Opticsmentioning

confidence: 99%

Adaptive optics: principles and applications in ophthalmology

Akyol

Hagag

Sivaprasad

et al. 2020

Eye

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This is a comprehensive review of the principles and applications of adaptive optics (AO) in ophthalmology. It has been combined with flood illumination ophthalmoscopy, scanning laser ophthalmoscopy, as well as optical coherence tomography to image photoreceptors, retinal pigment epithelium (RPE), retinal ganglion cells, lamina cribrosa and the retinal vasculature. In this review, we highlight the clinical studies that have utilised AO to understand disease mechanisms. However, there are some limitations to using AO in a clinical setting including the cost of running an AO imaging service, the time needed to scan patients, the lack of normative databases and the very small size of area imaged. However, it is undoubtedly an exceptional research tool that enables visualisation of the retina at a cellular level.

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“…When viewed directly, only light singly scattered from the point where the imaging beam is focused provides information about that location. Of course, fluorescence and multiply scattered light can also provide information when examined using specialized viewing methods based on intrinsic and extrinsic fluorophores and dark field microscopy (indirect viewing) (Chui et al 2012, Scoles et al 2014, Burns et al 2018, Hunter et al 2019, Jung et al 2019, Laforest et al 2020. While powerful, these methods do not detect single scatter, the focus of this review.…”

Section: Sensitivity (Weakest Reflectance To Be Detected)mentioning

confidence: 99%

Cellular-Scale Imaging of Transparent Retinal Structures and Processes Using Adaptive Optics Optical Coherence Tomography

Miller

Kurokawa

2020

Annu. Rev. Vis. Sci.

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High-resolution retinal imaging is revolutionizing how scientists and clinicians study the retina on the cellular scale. Its exquisite sensitivity enables time-lapse optical biopsies that capture minute changes in the structure and physiological processes of cells in the living eye. This information is increasingly used to detect disease onset and monitor disease progression during early stages, raising the possibility of personalized eye care. Powerful high-resolution imaging tools have been in development for more than two decades; one that has garnered considerable interest in recent years is optical coherence tomography enhanced with adaptive optics. State-of-the-art adaptive optics optical coherence tomography (AO-OCT) makes it possible to visualize even highly transparent cells and measure some of their internal processes at all depths within the retina, permitting reconstruction of a 3D view of the living microscopic retina. In this review, we report current AO-OCT performance and its success in visualizing and quantifying these once-invisible cells in human eyes. Expected final online publication date for the Annual Review of Vision Science, Volume 6 is September 15, 2020. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates.

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Longitudinal adaptive optics fluorescence microscopy reveals cellular mosaicism in patients

Cited by 26 publications

References 50 publications

Intravenous indocyanine green dye is insufficient for robust immune cell labelling in the human retina

Intravenous indocyanine green dye is insufficient for robust immune cell labelling in the human retina

Adaptive optics: principles and applications in ophthalmology

Cellular-Scale Imaging of Transparent Retinal Structures and Processes Using Adaptive Optics Optical Coherence Tomography

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