利用 Fiji 辅助的激光诱导脉络膜新生血管小鼠模型中脉络膜新生血管的自动定量容积分析。

Fiji-Assisted Automatic Quantitative Volumetric Analysis of Choroidal Neovascularization in a Laser-Induced Choroidal Neovascularization Mouse Model.

机构信息

USC Roski Eye Institute, Department of Ophthalmology, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA.

USC Ginsburg Institute for Biomedical Therapeutics, University of Southern California, Los Angeles, CA, USA.

出版信息

Transl Vis Sci Technol. 2023 Apr 3;12(4):10. doi: 10.1167/tvst.12.4.10.

DOI:10.1167/tvst.12.4.10

PMID:37043336

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10103716/

Abstract

PURPOSE

The laser-induced choroidal neovascularization (CNV) mouse model is the most frequently used animal model of CNV. To test new therapeutic agents that suppress CNV, CNV measurement in an accurate, precise, and efficient manner is important. We present the utility of Fiji-assisted automatic volumetric quantification of CNV in comparison with two-dimensional CNV analyses.

METHODS

Laser-induced CNV was induced in C57BL/6J mice according to the established protocol. After CNV induction, mice were treated with intravitreal injection of either phosphate-buffered saline solution (PBS) or Aflibercept, an anti- vascular endothelial growth factor agent. One week after intravitreal injection treatment, retina pigment epithelium/choroid flat mounts were stained with rhodamine-conjugated Griffonia simplicifolia lectin B4. Z-stacks of the entire CNV lesion obtained using laser confocal microscopy were converted to binary stacks using Fiji for volumetric analysis. Data from volumetric analysis and multiple area analyses from z-stack projection, the maximum, blindly selected, and mean area were compared using Fiji.

RESULTS

Fiji-assisted automatic quantitative volumetric analysis of CNV was useful in detecting experimental outliers in laser-induced CNV genesis and provided accurate and precise measurements of total areas of CNV with a lower coefficient of variance (63%) than in multiple area analyses, including the z-stack projection, maximum, blindly selected, and mean areas (67%, 67%, 76%, and 69%, respectively). A lower coefficient of variance in volumetric analysis than in multiple area analyses resulted in increased statistical significance when comparing CNV lesions in PBS, and Aflibercept-treated groups; P = 0.004 in volumetric analysis versus P value range between 0.03 and 0.05 in multiple area analyses.

CONCLUSIONS

Fiji-assisted automatic quantitative volumetric analysis can be useful for accurate, precise, and efficient measurements of total areas of CNV.

TRANSLATIONAL RELEVANCE

Volumetric measurement for CNV lesions can be advantageous in verifying the efficacy of therapeutic agents in the laser-induced CNV mouse model.

摘要

目的

激光诱导脉络膜新生血管（CNV）小鼠模型是 CNV 最常用的动物模型。为了测试抑制 CNV 的新治疗药物，以准确、精确和高效的方式测量 CNV 非常重要。我们介绍了 Fiji 辅助的 CNV 自动容积定量分析与二维 CNV 分析相比的效用。

方法

根据既定方案在 C57BL/6J 小鼠中诱导激光诱导的 CNV。CNV 诱导后，通过玻璃体内注射磷酸盐缓冲液（PBS）或抗血管内皮生长因子药物 Aflibercept 对小鼠进行治疗。玻璃体内注射治疗后 1 周，用 rhodamine 缀合的 Griffonia simplicifolia lectin B4 对视网膜色素上皮/脉络膜扁平膜进行染色。使用激光共聚焦显微镜获得的整个 CNV 病变的 Z 堆叠使用 Fiji 转换为二进制堆叠，用于容积分析。使用 Fiji 比较容积分析和来自 z 堆叠投影的多个区域分析的数据，包括最大、盲目选择和平均区域。

结果

Fiji 辅助的 CNV 自动定量容积分析有助于检测激光诱导 CNV 发生中的实验异常值，并提供 CNV 总区域的准确和精确测量，其变异系数（63%）低于多个区域分析，包括 z 堆叠投影、最大、盲目选择和平均区域（分别为 67%、67%、76%和 69%）。与多个区域分析相比，容积分析中的变异系数较低导致在比较 PBS 和 Aflibercept 治疗组的 CNV 病变时统计学意义增加；P = 0.004 用于容积分析，而 P 值范围在 0.03 到 0.05 之间，用于多个区域分析。