Name: quantitative fundus autofluorescence for the evaluation of retinal diseases
Uploaded: 2016-03-11T14:00:00.000+00:00
Duration: 7 min 22 s
Description: Watch this Scientific Journal Video about Quantitative Fundus Autofluorescence for the Evaluation of Retinal Diseases at JoVE.com

我们要感谢我们的合作者，弗朗索瓦Delori，托马斯·伯克和Tobias Duncker。 研究支持: NIH / NEI R01 EY015520（RTS，JPG），并且从研究经费不受限制防盲（RTB）。

伦理学声明:就读于这些研究所有患者均按照医学在纽约大学法学院批准的机构审查委员会的监督，以便完成。 1.患者的选择和成像前期准备注意:需要以下物质:0.5％托眼用溶液，2.5％苯肾上腺素滴眼液，CSLO配备谱域光学相干断层扫描（SD-OCT）和内部荧光参考。 <ol><li>成像之前，根据制造商的说明适当的设置CSLO进行数据采集。 </li><li>安装在CSLO内部荧光参考。注意:内部荧光基准，从制造商购买的，被容纳在一个金属环，并直接放置在照相机镜头的后面。如果使用从其它制造商的CSLO，可能有不同的结构比其在所述此协议。 <ol><li>为了插入引用到CSLO，扭动镜头将其删除，旋开机器的金属环，并包含引用的新的金属环代替它。 注意:内部荧光基准是将QAF技术必不可少的，因为它允许校正在激光功率和检测器灵敏度/增益变化。 </li></ol></li><li>招募成像患者接受常规散瞳检查，并获得有关过去的眼部历史和可能与眼部表现的调查结果基本医疗条件的背景资料。 </li><li>扩张0.5％托和2.5％苯的学生。关键的一步:扩张小学生至少6毫米。注:这是光线通过不间断的本质，从而为眼底的最佳可视化和测量。 </li><li>成像前，正确定位患者的CSLO，用下巴搁在腮托，额头抵靠foreheað休息，外侧眼角与指标正确对齐。 </li></ol> 2.眼底病的基线成像<ol><li>首先，图像，以便集中相机在黄斑部，并获得大致对焦近红外反射光（IR）（820波长）眼底。 <ol><li>与患者适当地定位，开关控制面板红外成像模式的硬件环境，位置相机手动直到眼底处于全焦点，拍摄图像</li><li>调节控制盘为"IR-OCT"，即用谱域光学相干断层扫描（SD-OCT）结合红外成像以评估潜在的疾病黄斑上的设置。 </li><li>使用目前在成像窗口的导向正确定位华侨城眼底的红外图像。以实现最佳的SD-OCT质量，摄像机的位置，使得OCT图像是在它的成像窗口的顶部三分之一。获得至少一个水平线SC通过中央凹的并跨越整个摄像视野。 </li></ol></li></ol> 3.设置QAF成像<ol><li>使用"高速"图像采集。注:此设置可以实现更快速的图像采集，从而减少信号损失的风险，由于患者的运动和由虹膜或眼睑的光造成堵塞。 </li><li>使用"平均9"帧。注:此设置允许9的图像帧​​，随后可以快速，连续捕获"平均"，以减少噪声和假象（见下文）。 </li><li>使用"30×30度"字段。注:这是指度图像采集过程中捕捉的视网膜区域。 </li><li>此前成像，警告蓝光耐心，因为这可以在第一次是惊人。 </li><li>打开自动对焦模式和对准照相机轴线，使得屏幕最大是"充满"与眼底AF（两侧和图像的角部的最小变暗）。 <ol><li>如果患者^ h容忍明亮的蓝色光大街的困难，开始用相机成像进一步远离眼睛，然后把相机缓慢地向患者直到眼底是在众目睽睽之下。 关键的一步:如果光线进入，或从眼排出受阻，降低信号将导致。局部减小是作为图像边或角的不对称变黑可见。当眼球运动单独阻碍光线通道信号的广义损失看到。 </li></ol></li><li>将照相机使得AF信号在整个领域的最高水平。瞄准最大的信号，而不是清晰的图像，但他们大致相关。通过移动CSLO手动或用检眼镜操纵杆重新调整相机调整相机焦距。 </li><li>调整灵敏度/增益，使得眼底AF是很容易看到，但要避免过度饱和。期间的图像采集，无论是在内部参考可见有色像素（位于的顶图像）或眼底表明过饱和并因此损失信号。 </li></ol> 4.图像采集注:关键的一步:在图像采集的目标应该是获得每个会话2高品质的9帧图像堆栈来控制一个会话内图像之间的差异。重新定位病人和相机后，获得两个图像的可变性一个第二次会议，评估和控制。所有图像将最终被校准到内部参考（以下描述）。 <ol><li>只有"漂白"（视网膜为AF的光的充分曝光）在感官视网膜9视紫红质，以尽量减少光的吸收的至少20秒后获得的图像。 <ol><li>利用这段时间来优化校准摄像机，对焦和感光度。 </li></ol></li><li>有一个新鲜的泪膜提高了信号质量，每个图像采集前闪烁的主题。 </li><li>避免在收购的平面上眼睑。 注意:它可能有助于手动打开眼皮更具挑战性的患者。笔者建议，助理执行此任务。 </li><li>每个图像采集之前优化对准，以确保光不被导致下降信号虹膜阻碍。 注:由于至少有轻微的运动是很常见的，作者建议每个图像采集前，根据需要精细调整。引导和鼓励患者，而成像有助于减少运动。作者还建议使用本机的脚踏板的"收购按钮"，以尽量减少操作者分心。 </li><li>通过计算"平均"的9帧堆栈，以增加信噪比进行图像后处理。计算使用CSLO软件，通过选择以计算平均值的期权的堆叠的平均值。 注:可能有堆栈，其中一些帧质量最优的不是（ 即有局限性或泛SI下降相对于最佳帧gnal），因此请仔细检查每个堆栈和平均计算之前取消选择次优的帧。 注意:具有可接受的信号 - 噪声比平均图像可以从至少3帧进行处理。这是正常的两侧和每个图像的四角比因CSLO本身的局限性中央20度较低的信号。 <ol><li>如果软件会询问经营者是否愿意正常化灰度级（ 即 ，舒展直方图）0和255之间，选择"否"。这保持了分析不变灰度级。 </li></ol></li></ol> 5.图像分析<ol><li>如以前使用的QAF技术开发的图像分析软件分析描述图像的自动对焦（IGOR; 9）。使用该软件的典型图像分析的简要说明包含下面。 <ol><li>加载从出口中IGOR的QAF程序，并导入图像（.bmp格式）CSLO软件的像素宽高比768 x 768（默认值输出参数）。 </li><li>选择下拉菜单中的影像，并开始分析。 </li><li>重新标定指标在图像上这样的"十字线"覆盖中心凹和"支架"紧靠视盘（这些指标被用于比例和位置的投资回报）​​。当正确定位时，使用软件提示然后填充周围中央凹圆周感兴趣区（ 见图1，2）的图像。 </li><li>选择在IGOR选项以每个图像的整体的灰度级校准中存在的内部荧光基准那些水平。注意:此步骤允许的机器有关的因素，包括电子零电平的各图像的各机器的内部参考校准因数，以及病人有关的因素，包括年龄，折射和角膜曲率校准。 注:分析软件然后显示固定ř所得到的图像和QAF值上egions论证每个区域内。 QAF值也自动输入到在单独的窗口的电子表格格式"。 </li><li>通过选择命令提示符中的选项生成QAF"热图"。所有的图像和数据可以从IGOR软件在Excel电子表格中选择相应的选项导出。 </li></ol></li></ol>

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STA: no disclosures, JPG: no disclosures, RTS: consultant, Ocata Therapeutics.

异常RPE脂褐素分布，是否增加或减少，是视网膜疾病的敏感标记物和一般与感官视网膜功能的丧失相关。在这里，我们描述了RPE脂褐素的评价QAF的应用。内部荧光参考掺入纠正可变激光功率和探测器的灵敏度9与我们的标准化的成像技术使得AF水平的可靠的定量。这是我们的目标，该方法将在诊断和视网膜疾病的监测帮助，并最终在评估治疗性干预，如药物或基因疗法的功效。 QAF也可协助?...

视网膜色素上皮细胞（RPE）支持感官视网膜通过众多方法1的功能。年龄相关性黄斑变性（AMD）是在工业化国家无法治疗失明的最重要的原因，其特征是在RPE中的变化，包括颜料的损失，功能和萎缩损失。在AMD和在正常的老化中，RPE积聚含有吞噬感光体的片段，称为脂褐素颗粒的荧光，溶酶体衍生的细胞器。 RPE脂褐素的累积已经认为表明氧化功能低下1，但最近的研究表明，视网膜色素上皮形态学老年眼睛保持正常高脂褐素水平2。然而，脂褐质分布异常模式，脂褐素的具体损失，都记录标志物AMD和AMD的进展，无论是组织学和临床3,4 缺陷PROCESRPE脂褐素唱也已显示在某些遗传性视网膜变性发生。患有斯特格病（STGD）遭受聚积在RPE脂褐素在年轻的时候，最终建立类似于AMD 5所示视力丧失。这些结果表明，脂褐质积聚本身可能是有毒的和驱动RPE功能障碍6,7。然而，受试者STGD随着时间的推移了详细的影像学研究并未证实焦点脂褐素的积累导致了随后的RPE损失8。因此，尽管脂褐质的异常是视网膜变性的标记，对于脂褐素的直接毒性作用尚未得到证实。 视网膜色素上皮是视网膜的最后的细胞层，但是产生从眼底的多数荧光信号。可使用共焦扫描激光眼底镜（CSLO），其允许六执行从RPE衍生产生和自体荧光（AF）的检测眼底AF的空间分布sualization。某些视网膜变性表明在这些条件下，诊断和监测眼底房颤独特的模式，和AF成像助剂。虽然标准的自动对焦成像是临床上重要的，定量的AF（QAF）已成为评估RPE健康的一个重要手段。我们和其他人开发了一种标准化的方法，能够可靠地确定在特定的视网膜部位9 QAF水平。 QAF在诊断和潜在的应用的视网膜条件监视，并且还可以具有在预后和风险分层效用。此外，QAF的诊断功能也已为某些视网膜疾病10-12所述。在这里，我们提供了对我们的技术伴随着它在健康和患病的眼睛的评价中的应用的可视化演示逐步细节。

<table><tbody><tr><td>Spectralis HRA + OCT</td><td>Heidelberg Engineering</td><td></td><td></td></tr><tr><td>0.5% tropicamide ophthalmic solution</td><td></td><td></td><td>Any brand can be used</td></tr><tr><td>2.5% phenylephrine ophthalmic solution</td><td></td><td></td><td>Any brand can be used</td></tr><tr><td>Internal fluorescent reference</td><td>Heidelberg Engineering</td><td></td><td></td></tr><tr><td>IGOR Pro software</td><td>WaveMetrics</td><td></td><td></td></tr></tbody></table>

quantitative fundus autofluorescence for the evaluation of retinal diseases

The retinal pigment epithelium (RPE) is juxtaposed to the overlying sensory retina, and supports the function of the visual system. Among the tasks performed by the RPE are phagocytosis and processing of outer photoreceptor segments through lysosome-derived organelles. These degradation products, stored and referred to as lipofuscin granules, are composed partially of bisretinoids, which have broad fluorescence absorption and emission spectra that can be detected clinically as fundus autofluorescence with confocal scanning laser ophthalmoscopy (cSLO). Lipofuscin accumulation is associated with increasing age, but is also found in various patterns in both acquired and inherited degenerative diseases of the retina. Thus, studying its pattern of accumulation and correlating such patterns with changes in the overlying sensory retina are essential to understanding the pathophysiology and progression of retinal disease. Here, we describe a technique employed by our lab and others that uses cSLO in order to quantify the level of RPE lipofuscin in both healthy and diseased eyes.

采用此技术，研究QAF在健康13和疾病状态10-12。在健康的眼睛（ 图1），从RPE发射AF时相对均匀地分布在整个眼底（ 图1A）。减少强度被认为是在黄斑中心区域由于黄斑色素阻挡光，并在两侧和图像的角由于眼睛和照相机的光学系统。血管暗，并应在明确的重点公采集的图像。 图1B说明了从图1A的QAF水平的相...

Watch this Scientific Journal Video about Quantitative Fundus Autofluorescence for the Evaluation of Retinal Diseases at JoVE.com

Quantitative Fundus Autofluorescence for the Evaluation of Retinal Diseases

定量自体荧光眼底视网膜疾病的评价

The retinal pigment epithelium (RPE) supports the sensory retina through recycling visual cycle byproducts, which accumulate as lipofuscin. These products are autofluorescent and can be qualitatively imaged in vivo. Here, we describe a method to quantitatively image RPE lipofuscin using confocal scanning laser ophthalmoscopy.

The retinal pigment epithelium (RPE) is juxtaposed to the overlying sensory retina, and supports the function of the visual system. Among the tasks ...

quantitative-fundus-autofluorescence-for-evaluation-retinal

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

11.4K 次观看.  NYU School of Medicine. The retinal pigment epithelium (RPE) supports the sensory retina through recycling visual cycle byproducts, which accumulate as lipofuscin. These products are autofluorescent and can be qualitatively imaged in vivo. Here, we describe a method to quantitatively image RPE lipofuscin using confocal scanning laser ophthalmoscopy. 

视频: Quantitative Fundus Autofluorescence for the Evaluation of Retinal Diseases

A Protocol to Evaluate and Quantify Retinal Pigmented Epithelium Pathologies in Mouse Models of Age-Related Macular Degeneration

Age-related macular degeneration (AMD) is a debilitating retinal disorder in aging populations. It is widely believed that dysfunction of the retinal pigmented epithelium (RPE) is a key pathobiological event in AMD. To understand the mechanisms that lead to RPE dysfunction, mouse models can be utilized by researchers. It has been established by previous studies that mice can develop RPE pathologies, some of which are observed in the eyes of individuals diagnosed with AMD. Here, we describe a phenotyping protocol to assess RPE pathologies in mice. This protocol includes the preparation and evaluation of retinal cross-sections using light microscopy and transmission electron microscopy, as well as that of RPE flat mounts by confocal microscopy. We detail the common types of murine RPE pathologies observed by these techniques and ways to quantify them through unbiased methods for statistical testing. As proof of concept, we use this RPE phenotyping protocol to quantify the RPE pathologies observed in mice overexpressing transmembrane protein 135 (Tmem135) and aged wild-type C57BL/6J mice. The main goal of this protocol is to present standard RPE phenotyping methods with unbiased quantitative assessments for scientists using mouse models of AMD.

Mouse models can be useful tools for investigating the biology of the retinal pigmented epithelium (RPE). It has been established that mice can develop an array of RPE pathologies. Here, we describe a phenotyping protocol to elucidate and quantify RPE pathologies in mice using light, transmission electron, and confocal microscopy.

评估和量化年龄相关性黄斑变性小鼠模型中视网膜色素上皮病理的协议

Age-related macular degeneration (AMD) is a debilitating retinal disorder in aging populations. It is widely believed that dysfunction of the retinal ...

科研

遗传学

Fundus Photography as a Convenient Tool to Study Microvascular Responses to Cardiovascular Disease Risk Factors in Epidemiological Studies

The microcirculation consists of blood vessels with diameters less than 150 &#181;m. It makes up a large part of the circulatory system and plays an important role in maintaining cardiovascular health. The retina is a tissue that lines the interior of the eye and it is the only tissue that allows for a non-invasive analysis of the microvasculature. Nowadays, high-quality fundus images can be acquired using digital cameras. Retinal images can be collected in 5 min or less, even without dilatation of the pupils. This unobtrusive and fast procedure for visualizing the microcirculation is attractive to apply in epidemiological studies and to monitor cardiovascular health from early age up to old age.
Systemic diseases that affect the circulation can result in progressive morphological changes in the retinal vasculature. For example, changes in the vessel calibers of retinal arteries and veins have been associated with hypertension, atherosclerosis, and increased risk of stroke and myocardial infarction. The vessel widths are derived using image analysis software and the width of the six largest arteries and veins are summarized in the Central Retinal Arteriolar Equivalent (CRAE) and the Central Retinal Venular Equivalent (CRVE). The latter features have been shown useful to study the impact of modifiable lifestyle and environmental cardiovascular disease risk factors.
The procedures to acquire fundus images and the analysis steps to obtain CRAE and CRVE are described. Coefficients of variation of repeated measures of CRAE and CRVE are less than 2% and within-rater reliability is very high. Using a panel study, the rapid response of the retinal vessel calibers to short-term changes in particulate air pollution, a known risk factor for cardiovascular mortality and morbidity, is reported. In conclusion, retinal imaging is proposed as a convenient and instrumental tool for epidemiological studies to study microvascular responses to cardiovascular disease risk factors.

Retinal image analysis is an unobtrusive procedure for visualizing the microcirculation. The impact of cardiovascular disease risk factors can result in changes of retinal vessel calibers. The procedures to acquire fundus images and the steps for calculating the vessel calibers are described.

眼底摄影作为一种方便的工具，研究微血管回应流行病学研究心血管疾病的风险因素

The microcirculation consists of blood vessels with diameters less than 150 &#181;m. It makes up a large part of the circulatory system and plays an ...

医学

Using Retinal Imaging to Study Dementia

The retina offers a unique &#8220;window&#8221; to study pathophysiological processes of dementia in the brain, as it is an extension of the central nervous system (CNS) and shares prominent similarities with the brain in terms of embryological origin, anatomical features and physiological properties.&#160; The vascular and neuronal structure in the retina can now be visualized easily and non-invasively using retinal imaging techniques, including fundus photography and optical coherence tomography (OCT), and quantified semi-automatically using computer-assisted analysis programs. Studying the associations between vascular and neuronal changes in the retina and dementia could improve our understanding of dementia and, potentially, aid in diagnosis and risk assessment.&#160; This protocol aims to describe a method of quantifying and analyzing retinal vasculature and neuronal structure, which are potentially associated with dementia. This protocol also provides examples of retinal changes in subjects with dementia, and discusses technical issues and current limitations of retinal imaging.

The retina shares prominent similarities with the brain and thus represents a unique window to study vasculature and neuronal structure in the brain non-invasively. This protocol describes a method to study dementia using retinal imaging techniques. This method can potentially aid in diagnosis and risk assessment of dementia.

利用视网膜成像研究痴呆

The retina offers a unique &#8220;window&#8221; to study pathophysiological processes of dementia in the brain, as it is an extension of the central ...

Experimental Autoimmune Uveitis: An Intraocular Inflammatory Mouse Model

Experimental Autoimmune Uveitis (EAU) is driven by immune cells responding to self-antigens. Many features of this non-infectious, intraocular inflammatory disease model recapitulate the clinical phenotype of posterior uveitis affecting humans. EAU has been used reliably to study the efficacy of novel inflammatory therapeutics, their mode of action and to further investigate the mechanisms that underpin disease progression of intraocular disorders. Here, we provide a detailed protocol on EAU induction in the C57BL/6J mouse - the most widely used model organism with susceptibility to this disease. Clinical assessment of disease severity and progression will be demonstrated using fundoscopy, histological examination and fluorescein angiography. The induction procedure involves subcutaneous&#160;injection of an emulsion containing a peptide (IRBP1-20) from the ocular protein interphotoreceptor retinoid binding protein (also known as retinol binding protein 3), Complete Freund&#39;s Adjuvant (CFA) and supplemented with killed Mycobacterium tuberculosis. Injection of this viscous emulsion on the back of the neck is followed by a single intraperitoneal injection of Bordetella pertussis toxin. At the onset of symptoms (day 12-14) and under general anesthesia, fundoscopic images are taken to assess disease progression through clinical examination. These data can be directly compared with those at later timepoints and peak disease (day 20-22) with differences analyzed. At the same time, this protocol allows the investigator to assess potential differences in vessel permeability and damage using fluorescein angiography. EAU can be induced in other mouse strains - both wildtype or genetically modified - and combined with novel therapies offering flexibility for studying drug efficacy and/or disease mechanisms.

In this report we present a protocol that allows the investigator to generate a mouse model of intraocular uveitis. More commonly referred to as experimental autoimmune uveitis (EAU), this established model captures many aspects of human disease. Herein, we will describe how to induce and monitor disease progression using several readouts.

实验性自身免疫性葡萄膜炎:眼内炎症小鼠模型

Experimental Autoimmune Uveitis (EAU) is driven by immune cells responding to self-antigens. Many features of this non-infectious, intraocular ...

免疫与感染

用于 AMD 研究的基于 OCT 的人类供体眼多模态成像

Ex Vivo OCT-Based Multimodal Imaging of Human Donor Eyes for Research into Age-Related Macular Degeneration

A progression sequence for age-related macular degeneration (AMD) learned from optical coherence tomography (OCT)-based multimodal (MMI) clinical imaging could add prognostic value to laboratory findings. In this work, ex vivo OCT and MMI were applied to human donor eyes prior to retinal tissue sectioning. The eyes were recovered from non-diabetic white donors aged &#8805;80 years old, with a death-to-preservation time (DtoP) of &#8804;6 h. The globes were recovered on-site, scored with an 18 mm trephine to facilitate cornea removal, and immersed in buffered 4% paraformaldehyde. Color fundus images were acquired after anterior segment removal with a dissecting scope and an SLR camera using trans-, epi-, and flash illumination at three magnifications. The globes were placed in a buffer within a custom-designed chamber with a 60 diopter lens. They were imaged with spectral domain OCT (30&#176; macula cube, 30 &#181;m spacing, averaging = 25), near-infrared reflectance, 488 nm autofluorescence, and 787 nm autofluorescence. The AMD eyes showed a change in the retinal pigment epithelium (RPE), with drusen or subretinal drusenoid deposits (SDDs), with or without neovascularization, and without evidence of other causes. Between June 2016 and September 2017, 94 right eyes and 90 left eyes were recovered (DtoP: 3.9 &plusmn; 1.0 h). Of the 184 eyes, 40.2% had AMD, including early intermediate (22.8%), atrophic (7.6%), and neovascular (9.8%) AMD, and 39.7% had unremarkable maculas. Drusen, SDDs, hyper-reflective foci, atrophy, and fibrovascular scars were identified using OCT. Artifacts included tissue opacification, detachments (bacillary, retinal, RPE, choroidal), foveal cystic change, an undulating RPE, and mechanical damage. To guide the cryo-sectioning, OCT volumes were used to find the fovea and optic nerve head landmarks and specific pathologies. The ex vivo volumes were registered with the in vivo volumes by selecting the reference function for eye tracking. The ex vivo visibility of the pathology seen in vivo depends on the preservation quality. Within 16 months, 75 rapid DtoP donor eyes at all stages of AMD were recovered and staged using clinical MMI methods.

作者聚焦:基于离体 OCT 的人类供体眼多模态成像，用于年龄相关性黄斑变性的研究  

Laboratory assays can leverage prognostic value from the longitudinal optical coherence tomography (OCT)-based multimodal imaging of age-related macular degeneration (AMD). Human donor eyes with and without AMD are imaged using OCT, color, near-infrared reflectance scanning laser ophthalmoscopy, and autofluorescence at two excitation wavelengths prior to tissue sectioning.

离体 基于OCT的人供体眼睛多模态成像用于研究年龄相关性黄斑变性

A progression sequence for age-related macular degeneration (AMD) learned from optical coherence tomography (OCT)-based multimodal (MMI) clinical imaging ...

神经科学

Retinal Pathophysiological Evaluation in a Rat Model

A posterior segment eye disease like diabetic retinopathy alters the physiology of the retina. Diabetic retinopathy is characterized by a retinal detachment, breakdown of the blood-retinal barrier (BRB), and retinal angiogenesis. An in vivo rat model is a valuable experimental tool to examine the changes in the structure and function of the retina. We propose three different experimental techniques in the rat model to identify morphological changes of retinal cells, retinal vasculature, and compromised BRB. Retinal histology is used to study the morphology of various retinal cells. Also, quantitative measurement is performed by retinal cell count and thickness measurement of different retinal layers. A BRB breakdown assay is used to determine the leakage of extraocular proteins from the plasma to vitreous tissue due to the breakdown of BRB. Fluorescence angiography is used to study angiogenesis and leakage of blood vessels by visualizing retinal vasculature using FITC-dextran dye.

Diabetic retinopathy is one of the leading causes of blindness. Histology, blood-retinal barrier breakdown assay, and fluorescence angiography are valuable techniques to understand the pathophysiology of the retina, which could further enhance the efficient drug screening against diabetic retinopathy.

大鼠模型中的视网膜病理生理学评估

A posterior segment eye disease like diabetic retinopathy alters the physiology of the retina. Diabetic retinopathy is characterized by a retinal ...

Image-Guided Optical Coherence Tomography to Assess Structural Changes in Rodent Retinas

Ocular diseases, such as age-related macular degeneration, glaucoma, retinitis pigmentosa, and uveitis, are always accompanied by retinal structural changes. These diseases affecting the fundus always exhibit typical abnormalities in certain cell types in the retina, including photoreceptor cells, retinal ganglion cells, cells in the retinal blood vessels, and cells in the choroidal vascular cells. Noninvasive, highly efficient, and adaptable imaging techniques are required for both clinical practice and basic research. Image-guided optical coherence tomography (OCT) satisfies these requirements because it combines fundus photography and high-resolution OCT, providing an accurate diagnosis of tiny lesions as well as important changes in the retinal architecture. This study details the procedures of data collection and data analysis for image-guided OCT and demonstrates its application in rodent models of choroidal neovascularization (CNV), optic nerve crush (ONC), light-induced retinal degeneration, and experimental autoimmune uveitis (EAU). This technique helps researchers in the eye field to identify rodent retinal structural changes conveniently, reliably, and tractably.

The protocol presented here details the procedures of data collection and data analysis for image-guided optical coherence tomography (OCT) and demonstrates its application in multiple rodent models of ocular diseases.

Ocular diseases, such as age-related macular degeneration, glaucoma, retinitis pigmentosa, and uveitis, are always accompanied by retinal structural ...

探索年龄相关性黄斑变性中的定量自发荧光

A Workflow to Quantitatively Determine Age-Related Macular Degeneration Lesion-Specific Variations in Fundus Autofluorescence

Fundus autofluorescence (FAF) imaging allows the noninvasive mapping of intrinsic fluorophores of the ocular fundus, particularly the retinal pigment epithelium (RPE), now quantifiable with the advent of confocal scanning laser ophthalmoscopy-based quantitative autofluorescence (QAF). QAF has been shown to be generally decreased at the posterior pole in age-related macular degeneration (AMD). The relationship between QAF and various AMD lesions (drusen, subretinal drusenoid deposits) is still unclear.
This paper describes a workflow to determine lesion-specific QAF in AMD. A multimodal in vivo imaging approach is used, including but not limited to spectral domain optical coherence tomography (SD-OCT) macular volume scanning and QAF. Using customized FIJI plug-ins, the corresponding QAF image is aligned with the near-infrared image from the SD-OCT scan (characteristic landmarks; i.e., vessel bifurcations). The foveola and the edge of the optic nerve head are marked in the OCT images (and transferred to the registered QAF image) for accurate positioning of the analysis grids.
AMD-specific lesions can then be marked on individual OCT BScans or the QAF image itself. Normative QAF maps are created to account for the varying mean and standard deviation of QAF values throughout the fundus (QAF images from a representative AMD group were averaged to build normative standard retinal QAF AMD maps). The plug-ins record the X and Y coordinates, z-score (a numerical measurement that describes the QAF value in relation to the mean of AF maps in terms of standard deviation from the mean), mean intensity value, standard deviation, and number of pixels marked. The tools also determine z-scores from the border zone of marked lesions.&#160;This workflow and the analysis tools will improve the understanding of the pathophysiology and clinical AF image interpretation in AMD.

作者聚焦:使用 QAF 工作流程了解年龄相关性黄斑变性的病理生理学 

This research describes a workflow to determine and compare autofluorescence levels from individual regions of interest (e.g., drusen and subretinal drusenoid deposits in age-related macular degeneration [AMD]) while accounting for varying autofluorescence levels throughout the fundus.

定量确定年龄相关性黄斑变性病变性病变性眼底自发荧光特异性变化的工作流程

Fundus autofluorescence (FAF) imaging allows the noninvasive mapping of intrinsic fluorophores of the ocular fundus, particularly the retinal pigment ...

定量自体荧光眼底视网膜疾病的评价

摘要

探索更多视频

此视频中的章节

评估和量化年龄相关性黄斑变性小鼠模型中视网膜色素上皮病理的协议

眼底摄影作为一种方便的工具，研究微血管回应流行病学研究心血管疾病的风险因素

利用视网膜成像研究痴呆

实验性自身免疫性葡萄膜炎:眼内炎症小鼠模型

离体基于OCT的人供体眼睛多模态成像用于研究年龄相关性黄斑变性

离体基于OCT的人供体眼睛多模态成像用于研究年龄相关性黄斑变性

离体基于OCT的人供体眼睛多模态成像用于研究年龄相关性黄斑变性

大鼠模型中的视网膜病理生理学评估

Image-Guided Optical Coherence Tomography to Assess Structural Changes in Rodent Retinas

定量确定年龄相关性黄斑变性病变性病变性眼底自发荧光特异性变化的工作流程

定量自体荧光眼底视网膜疾病的评价

摘要

探索更多视频

此视频中的章节

评估和量化年龄相关性黄斑变性小鼠模型中视网膜色素上皮病理的协议

眼底摄影作为一种方便的工具，研究微血管回应流行病学研究心血管疾病的风险因素

利用视网膜成像研究痴呆

实验性自身免疫性葡萄膜炎:眼内炎症小鼠模型

离体 基于OCT的人供体眼睛多模态成像用于研究年龄相关性黄斑变性

离体 基于OCT的人供体眼睛多模态成像用于研究年龄相关性黄斑变性

离体 基于OCT的人供体眼睛多模态成像用于研究年龄相关性黄斑变性

大鼠模型中的视网膜病理生理学评估

Image-Guided Optical Coherence Tomography to Assess Structural Changes in Rodent Retinas

定量确定年龄相关性黄斑变性病变性病变性眼底自发荧光特异性变化的工作流程

离体基于OCT的人供体眼睛多模态成像用于研究年龄相关性黄斑变性

离体基于OCT的人供体眼睛多模态成像用于研究年龄相关性黄斑变性

离体基于OCT的人供体眼睛多模态成像用于研究年龄相关性黄斑变性