研究目的
To investigate a multimodal imaging system for monitoring retinal neovascularization using PAM, OCT, FA, and color fundus photography in living rabbits.
研究成果
The multimodal SD-OCT and OR-PAM imaging system effectively detects and visualizes retinal neovascularization in vivo with high resolution and contrast, using safe laser energy levels. It provides 3D volumetric images and quantitative data on vascular changes, offering a non-invasive, label-free method for ophthalmic diagnosis and monitoring.
研究不足
The acquisition time for PAM and OCT is longer compared to FA and color fundus photography, limiting real-time monitoring. The study was conducted in rabbits, and translation to human clinical applications requires further safety evaluations for long-term effects of laser exposure. The RNV model shows dynamic changes over time, which may not fully mimic human disease progression.
1:Experimental Design and Method Selection:
A custom-built multimodal imaging system combining optical-resolution photoacoustic microscopy (OR-PAM) and spectral domain optical coherence tomography (SD-OCT) was used. The system was designed to provide high-resolution, non-invasive imaging of retinal vasculature based on optical absorption and back-scattering properties.
2:Sample Selection and Data Sources:
Eight New Zealand rabbits were used, with retinal neovascularization induced by intravitreal injection of VEGF-
3:Imaging was performed at multiple time points (days 0, 4, 5, 6, 7, 9, 11, 14, 28, 35 post-injection). List of Experimental Equipment and Materials:
1 Equipment included a custom-built dual PAM and OCT system, Topcon 50EX fundus camera, ultrasound transducer, laser sources, galvanometer scanner, and various chemicals (VEGF-165, PBS, fluorescein, anesthetics). Materials were purchased from specified suppliers.
4:Experimental Procedures and Operational Workflow:
Rabbits were anesthetized, pupils dilated, and VEGF injected. Imaging sessions involved sequential acquisition of color fundus, FA, OCT, and PAM images. PAM was performed at 580 nm wavelength with laser energy below ANSI safety limits. Data were processed using Amira software for 3D reconstruction and segmentation.
5:Data Analysis Methods:
Quantitative analysis of vessel density and diameter was done using ImageJ software. Statistical analysis used Student's t-tests with p ≤ 0.05 significance level. Histological analysis with H&E staining was performed post-sacrifice.
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Ganymede-II-HR
Ganymede-II-HR
Thorlabs
Spectral domain optical coherence tomography system for high-resolution retinal imaging.
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Scan Lens
OCT-LK3-BB
Thorlabs
Part of the telescope in the imaging system to focus light on the fundus.
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Ophthalmic Lens
AC080-010-B-ML
Thorlabs
Focuses the scanned beam on the fundus using the rabbit eye optics.
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Topcon 50EX
50EX
Topcon Corporation
Used for color fundus photography and fluorescein angiography to monitor retinal vessels.
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Ultrasound Transducer
27.0 MHz needle-shaped
Optosonic Inc.
Detects laser-induced acoustic signals in the photoacoustic microscopy system.
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Digitizer
PX1500-4
Signatec Inc.
High-speed digitizer for sampling photoacoustic signals.
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Amira Software
FEI
Used for 3D image reconstruction and segmentation of retinal vasculature.
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ImageJ Software
National Institute of Health
Used for quantitative analysis of vessel density and diameter.
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Leica DM600 Light Microscope
DM600
Leica Biosystems
Used for histological examination and imaging of tissue sections.
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Pulse Oximeter
V8400D Capnograph & SpO2 Digital Pulse Oximetry
Smiths Medical
Monitors heart rate and respiratory rate of rabbits during experiments.
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Water-circulating Blanket
TP-700
Stryker Corporation
Maintains animal body temperature during experiments.
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