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Updated: Jul 6, 2026
In Vivo Multimodal Imaging and Analysis of Mouse Laser-Induced Choroidal Neovascularization Model
Published on: January 21, 2018
Effects of an anti-VEGF-A monoclonal antibody on laser-induced choroidal neovascularization in mice: optimizing
Claudio Campa1, Ian Kasman, Weilan Ye
1Genentech, Inc., South San Francisco, California, USA. claudio.campa@yahoo.com
Purpose:
The purpose of this study was to evaluate different methods of detecting and quantifying experimentally induced choroidal neovascularization (CNV) and vascular changes induced on CNV by an anti-VEGF-A monoclonal antibody.
Methods:
Choroidal neovascularization was induced by 532-nm diode laser in C57BL/6 mice. Ten days after the laser, the following methods were used to detect the new vessels: high-resolution angiography with fluorescein isothiocyanate-dextran; immunohistochemistry with biotinylated isolectin, rabbit anti-NG2, rat anti-CD31, rabbit anti-VWF, rat ani-CD105, rabbit anti-collagen IV, rat anti-ICAM-2, rabbit anti-desmin, and rat anti-MECA 32; and intravital injection of fluorescein-labeled Lycopersicon esculentum (tomato) lectin. To verify the validity of these staining methods in the quantification of treated CNV, the authors applied the most effective of these techniques to three groups of mice after laser induction of CNV and treatment with an anti-VEGF full antibody (G6-31).
Results:
Fluorescein isothiocyanate-dextran angiography, rat anti-ICAM-2 immunostaining, and tomato lectin intravital injection resulted in the most effective means of identifying choroidal neovascularization. A certain amount of nonspecific fluorescence was detected in the area of CNV for each
Method:
This fluorescence appeared more intense when fluorescein isothiocyanate-dextran was used. Tomato lectin injection and rat anti-ICAM-2 immunostaining were the methods that better recorded the antiangiogenic drug effect.
Conclusions:
Because of easy execution, low background fluorescence, and detailed visualization of new vessels, intravital injection of tomato lectin followed by a quantification based on threshold fluorescence represents the best technique for measuring CNV and the vascular changes induced by anti-VEGF-A monoclonal antibody in mice.
Insights
Intravital injection of tomato lectin is the best method for detecting choroidal neovascularization (CNV) and anti-VEGF-A antibody effects in mice, offering easy execution and clear visualization.
Area of Science:
- Ophthalmology
- Angiogenesis Research
- Animal Models
Background:
- Choroidal neovascularization (CNV) is a key factor in vision loss.
- Accurate detection and quantification of CNV are crucial for evaluating treatments.
- Existing methods for CNV assessment have limitations.
Purpose of the Study:
- To evaluate various methods for detecting and quantifying laser-induced CNV in mice.
- To assess the effectiveness of these methods in visualizing vascular changes induced by an anti-VEGF-A antibody.
Main Methods:
- CNV was induced in C57BL/6 mice using a diode laser.
- Detection methods included high-resolution angiography, immunohistochemistry (targeting various markers like CD31, CD105, ICAM-2), and intravital tomato lectin injection.
- Effectiveness was validated by quantifying CNV after anti-VEGF-A antibody treatment.
Main Results:
- Fluorescein angiography, ICAM-2 immunostaining, and tomato lectin injection were most effective for identifying CNV.
- Tomato lectin and ICAM-2 methods better captured the antiangiogenic effects of the drug.
- Fluorescein isothiocyanate-dextran showed more intense, nonspecific fluorescence.
Conclusions:
- Intravital tomato lectin injection is the optimal technique for measuring CNV in mice.
- This method offers easy execution, minimal background fluorescence, and detailed visualization of neovascularization.
- It is highly effective for assessing vascular changes induced by anti-VEGF-A antibodies.
