Name: measuring retinal vessel diameter from mouse fluorescent angiography images
Uploaded: 2023-05-19T13:20:00
Description: Watch this Scientific Journal Video about Measuring Retinal Vessel Diameter from Mouse Fluorescent Angiography Images at JoVE.com

这项工作得到了冰岛研究基金（217796-052）（AGL）和HelgaJónsdóttir和Sigurlidi Kristjánsson纪念基金（AGL和TE）的博士后奖学金支持。作者感谢Eiríkur Steingrímsson教授提供小鼠。

所有实验均获得冰岛食品和兽医局的批准（MAST 许可证编号 2108002）。所有动物研究均根据视觉和眼科研究协会 （ARVO） 关于在眼科和视力研究中使用动物的声明进行。本研究使用雄性和雌性 C57BL/6J 和 Mitfmi-vga9/+ 小鼠。将C57BL / 6J小鼠（n = 7）用作对照。野生型是商业获得的（见 材料表），但所有突变小鼠（n = 7）都是在冰岛大学生物医学中心的动物设施中繁殖和饲养的?...

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本文首次提出了一种从小鼠FA图像中分析视网膜血管直径和视网膜血管系统的方法。由于仅使用眼底成像来捕获视网膜血管系统的图像，因此该方法有几个缺点，其中之一是只能推断出本研究中检查的小鼠视网膜血管系统浅层的变化;更深层次的任何差异都尚不清楚。 
已经提出了一种独特的光学相干断层扫描血管造影（OCTA）图像分析方法，该方法使用自动血管追踪和血管直径...

可能人体中研究最多的血管床是视网膜血管系统。随着技术的不断提高，视网膜血管系统很容易在活着的患者身上拍摄，并被用于许多研究领域1.此外，小鼠视网膜血管系统在发育过程中已被证明是研究血管生长基础生物学的非常有效的模型系统。视网膜血管系统的主要目的是通过渗透神经组织的层状毛细血管网为视网膜内部提供代谢支持2。然而，视网膜的状况，以及由此产生的任何功能障碍或萎缩，都会对视网膜血管系统的分叉和动脉直径产生显着影响，这表明视网膜细胞和血管系统之间存在相互作用3,4。众所周知，许多眼部疾病，包括早产儿视网膜病变 （ROP）、糖尿病视网膜病变 （DR）、年龄相关性黄斑变性 （AMD）、青光眼和角膜新生血管形成，可导致眼部血管生成异常5。在视网膜血管系统的情况下，视网膜变性的小鼠模型通常表现出与人类血管疾病中观察到的变化相当6,7。基本螺旋-环-螺旋-拉链转录因子的 Myc 超基因家族包括在视网膜色素上皮 （RPE） 中表达的小眼症相关转录因子 （Mitf） 基因8,9,10。
许多器官，包括眼睛、耳朵、免疫系统、中枢神经系统、肾脏、骨骼和皮肤，已被证明受 Mitf 9,11,12,13 的调节。 我们发现，在携带 Mitf 基因各种突变的小鼠中，RPE 的结构和功能会受到影响，导致一些视网膜变性并最终导致视力丧失10。最近，已经表明，Mitf突变型和野生型小鼠14之间的血管数量和血管直径存在显着差异。由于视网膜成像的发展，研究人员和医生现在可以精确量化体内的视网膜血管系统。自 1800 年代以来，研究人员和医生利用了视网膜血管系统可视化的好处，荧光素血管造影 （FA） 已显示视网膜血流和血液-视网膜屏障的降解15。
本文演示了如何使用 MATLAB 软件中的自定义编写代码分析小鼠 FA 图像中的视网膜血管直径。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>1% Tropicamide (Mydriacyl)</td>
			<td>Alcon Inc Laboratories</td>
			<td></td>
			<td>Mydriatic agent</td>
		</tr>
		<tr>
			<td>2% Methocel</td>
			<td>OmniVision Eye Care</td>
			<td></td>
			<td>Hydroxypropryl methylcellulose gel</td>
		</tr>
		<tr>
			<td>C57BL/6J</td>
			<td>Jackson Laboratory</td>
			<td>000664</td>
			<td>Wild type mice</td>
		</tr>
		<tr>
			<td>Chanazine 2% (xylazine)</td>
			<td>Chanelle Animal Health UK</td>
			<td>BN I21322/I</td>
			<td>Anesthesia IP</td>
		</tr>
		<tr>
			<td>Excel for Microsoft 365</td>
			<td>Microsoft Inc</td>
			<td></td>
			<td>Software package</td>
		</tr>
		<tr>
			<td>Fluorescein sodium salt</td>
			<td>Sigma-Aldrich</td>
			<td>28803-100G</td>
			<td>Fluorescent angiography</td>
		</tr>
		<tr>
			<td>Matlab 8.0</td>
			<td>The MathWorks, Inc.</td>
			<td></td>
			<td>Software package</td>
		</tr>
		<tr>
			<td>Micron IV rodent fundus camera</td>
			<td>Phoenix-Micron</td>
			<td>40-2200</td>
			<td>Fundus photography</td>
		</tr>
		<tr>
			<td>Phenylephrine 10% w/v</td>
			<td>Bausch &#38; Lomb</td>
			<td></td>
			<td>Mydriatic agent</td>
		</tr>
		<tr>
			<td>Phosphate Buffered Saline - 100 tablets</td>
			<td>Gibco</td>
			<td>18912-014</td>
			<td>Dilution</td>
		</tr>
		<tr>
			<td>Sigmaplot 13</td>
			<td>Jandel Scientific Software</td>
			<td></td>
			<td>Software package</td>
		</tr>
		<tr>
			<td>S-Ketamine, 25 mg/mL</td>
			<td>Pfizer Inc.</td>
			<td>PAA104470</td>
			<td>Anesthesia IP</td>
		</tr>
	</tbody>
</table>

measuring retinal vessel diameter from mouse fluorescent angiography images

研究视网膜病变中视网膜血管系统的发育非常重要，其中异常血管生长最终会导致视力丧失。小眼症相关转录因子 （Mitf） 基因突变表现为色素减退、小眼症、视网膜变性，在某些情况下还会出现失明。通过非侵入性手段对小鼠视网膜进行 体内 成像对于眼科研究至关重要。然而，鉴于其体积小，小鼠眼底成像很困难，可能需要专门的工具、维护和培训。在这项研究中，我们开发了一种独特的软件，可以使用MATLAB编写的自动化程序分析小鼠的视网膜血管直径。在腹膜内注射荧光素盐溶液后，使用商业眼底照相机系统获得眼底照片。对图像进行了更改以增强对比度，并且MATLAB程序允许在距视盘的预定距离处自动提取平均血管直径。通过分析视网膜血管直径，研究了野生型小鼠和 Mitf 基因不同突变小鼠的血管变化。这里开发的定制编写的MATLAB程序实用，易于使用，并允许研究人员方便可靠地分析小鼠视网膜脉管系统的平均直径和平均总直径，以及血管数量。

图 1 显示了用于分析视网膜血管系统的过程，该过程应用于所有测试小鼠的小鼠 FFA 图像。半径是光盘的两倍，用于测量从光盘中心开始沿顺时针方向旋转的像素强度。当它分别遇到高于和低于用户指定阈值的点时，它会用起点或终点标记像素。重复 30 次，每次都离视盘中心稍远一点。在荧光素注射后5分钟拍摄FFA图像（图1A 和 图1B</stro...

Watch this Scientific Journal Video about Measuring Retinal Vessel Diameter from Mouse Fluorescent Angiography Images at JoVE.com

Measuring Retinal Vessel Diameter from Mouse Fluorescent Angiography Images

小鼠视网膜血管系统在理解血管模式形成的机制方面特别有趣。该协议在距视盘固定距离处自动测量荧光血管造影眼底图像的小鼠视网膜血管直径。

从小鼠荧光血管造影图像中测量视网膜血管直径

It is important to study the development of retinal vasculature in retinopathies in which abnormal vessel growth can ultimately lead to vision loss. ...

该协议提供了一种简单可靠的方法，可以自动测量由于基因突变或视网膜疾病而麻醉小鼠的视网膜血管直径。这种技术的主要优点是这里定制编写的MATLAB程序实用、易用、方便、可靠。视网膜血管直径变化通常发生在退行性疾病中，如 RP 和其他视网膜血管疾病。小鼠模型提供了对治疗效果及其对视网膜血管影响的见解。该方法可能作为眼部血管疾病进展后的可量化临床标志物，以及监测可能的治疗方法。首先，准备一个装有荧光素工作溶液的注射器，并将其腹膜内给药给药给麻醉的动物，并在角膜表面滴一滴2%甲基纤维素凝胶。然后将动物放在成像系统的定位台上。将视网膜成像眼底相机镜头直接轻轻地触摸鼠标的角膜，并稍微调整对准方式，将视神经头置于视野的中间。然后切换到眼底相机上的绿色荧光通道，并聚焦在视网膜血管上以拍摄图像。在荧光素注射后的所需时间点捕获图像，以确定理想的时间点并可视化荧光素眼底图像。首先，打开MATLAB程序。然后下载并保存眼底直径。M 代码。接下来，打开眼底图像分析文件夹并拖放眼底直径。M 代码添加到分析文件夹中。将 FFA 图像拖放到图像分析文件夹中。在MATLAB程序中，在代码文件上单击两次以激活它。选择“运行”选项。将出现一个弹出菜单以选择图像。选择图像，屏幕上将出现一个用于分析参数的弹出菜单。在弹出菜单中设置分析参数，然后按 OK。选择光盘的中心，然后选择光盘的边缘。分析将在后台执行，分析结果将显示在屏幕上。该表包含每艘船的平均值、标准差和标准误差值。保存表。然后保存设置了平均厚度的图形。打开包含已保存文件的分析文件夹，然后打开已保存的图窗。打开保存的表，复制值，然后将它们粘贴到 Excel 文件中。荧光素注射后，获得来自对照小鼠和MITFMI-VGA9 /突变小鼠的图像，其中从视盘中心到血管端点的最短距离沿顺时针方向测量。平均血管直径是作为野生型和突变型小鼠的血管数量的函数获得的。要记住的最重要的事情是眼底摄像头的位置和视盘在眼底图像中的居中。除了这种技术外，还可以进行分子技术，以探索新药，诊断眼睛中的不同血管状况，并更好地了解视网膜血管系统的功能。

Research

JoVE Journal

Medicine

Detecting Abnormalities in Choroidal Vasculature in a Mouse Model of Age-related Macular Degeneration by Time-course Indocyanine Green Angiography

按时间当然吲哚菁绿血管造影检测脉络膜血管系统异常在年龄相关性黄斑变性的小鼠模型

Indocyanine Green&#160;Angiography (or ICGA) is a technique performed by ophthalmologists to diagnose abnormalities of the choroidal and retinal ...

科研

医学

Ferric Chloride-induced Thrombosis Mouse Model on Carotid Artery and Mesentery Vessel

三氯化铁诱发血栓形成小鼠模型上的颈动脉和肠系膜血管

Severe thrombosis and its ischemic consequences such as myocardial infarction, pulmonary embolism and stroke are major worldwide health issues. The ferric ...

Fluorescent Dye Labeling of Erythrocytes and Leukocytes for Studying the Flow Dynamics in Mouse Retinal Circulation

用于研究小鼠视网膜循环中流动动力学的红细胞和白细胞的荧光染料标记

The retinal and choroidal blood flow dynamics may provide insight into the pathophysiology and sequelae of various ocular diseases, such as glaucoma, ...

Preparation Steps for Measurement of Reactivity in Mouse Retinal Arterioles Ex Vivo

Preparation Steps for Measurement of Reactivity in Mouse Retinal Arterioles Ex Vivo

小鼠视网膜微动脉的反应性测定的制备步骤前体

Vascular insufficiency and alterations in normal retinal perfusion are among the major factors for the pathogenesis of various sight-threatening ocular ...

Vessel-sparing Excision and Primary Anastomosis

血管保留切除术与原发性吻合术

Urethroplasty is considered to be the standard treatment for urethral strictures since it provides excellent long-term success rates. For isolated short ...

Two-vessel Occlusion Mouse Model of Cerebral Ischemia-reperfusion

脑缺血再灌注的双血管闭塞小鼠模型

In this study, a middle cerebral artery (MCA) occlusion mouse model is employed to study cerebral ischemia-reperfusion. A reproducible and reliable mouse ...

Retinal Organoid Induction System for Derivation of 3D Retinal Tissues from Human Pluripotent Stem Cells

视网膜器官诱导系统，用于从人类多能干细胞中提取3D视网膜组织

Retinal degenerative diseases are the main causes of irreversible blindness without effective treatment. Pluripotent stem cells that have the potential to ...

Evaluation of Capillary and Other Vessel Contribution to Macular Perfusion Density Measured with Optical Coherence Tomography Angiography

评估毛细血管和其他血管对光学相干断层扫描血管造影测量的黄斑灌注密度的贡献

Parafoveal circulation of the superficial retinal capillary plexus is usually measured with vessel density, which determines the length of capillaries ...

Standardized Rat Coronary Ring Preparation and Real-Time Recording of Dynamic Tension Changes Along Vessel Diameter

标准化大鼠冠状动脉环制备及实时记录血管直径动态张力变化

As a key event of cardiovascular system diseases, coronary artery disease (CAD) has been widely regarded as the main culprit of atherosclerosis, ...

Retinal Explant of the Adult Mouse Retina as an Ex Vivo Model for Studying Retinal Neurovascular Diseases

Retinal Explant of the Adult Mouse Retina as an Ex Vivo Model for Studying Retinal Neurovascular Diseases

成年小鼠视网膜的视网膜外植体作为研究视网膜神经血管疾病的离 体 模型

One of the challenges in retina research is studying the cross-talk between different retinal cells such as retinal neurons, glial cells, and vascular ...