研究目的
To demonstrate that indocyanine green (ICG), an FDA-approved dye, can be used as a control agent in paired-agent imaging of sentinel lymph nodes (PAISLY) for antibody-based targeted agents by comparing its kinetics with fluorescently labeled antibodies in rat lymph nodes.
研究成果
ICG, when premixed with albumin, can effectively serve as a control agent in PAISLY for antibody-based targeted imaging agents, enabling non-invasive detection of lymph node metastases with high sensitivity. This facilitates clinical translation by utilizing FDA-approved components.
研究不足
The study is limited to rat models and may not directly translate to humans. The mixing protocol for ICG and albumin is critical and must be optimized to avoid issues like self-quenching and binding interference. The use of DMSO in ICG preparation, though minimal, could affect results compared to clinical formulations. The approach requires further validation with FDA-approved targeted antibodies for clinical use.
1:Experimental Design and Method Selection:
The study used paired-agent imaging (PAISLY) with compartmental modeling to compare the kinetics of ICG and antibody-based imaging agents in rat lymph nodes.
2:Sample Selection and Data Sources:
Female Fischer 344 rats (n=5) for control lymph nodes and female athymic nude rats (n=4) for metastatic lymph nodes were used. Cancer cells (MDA-MB-231-D3H2LN-GFP) were implanted in the footpad for metastasis model.
3:List of Experimental Equipment and Materials:
Equipment includes Pearl Impulse Small Animal Imaging System, cryostat microtome, fluorescence microscope, NanoDrop spectrophotometer, and MATLAB for analysis. Materials include ICG, IRDye-700DX, antibodies (IgG and cetuximab), albumin, DMSO, PBS, and anesthesia (isoflurane).
4:Experimental Procedures and Operational Workflow:
Imaging agents were prepared and co-injected intradermally into the footpad. Fluorescence images were acquired at 1-minute intervals for 2 hours. Data were analyzed using MATLAB for kinetic modeling and binding potential calculation.
5:Data Analysis Methods:
Statistical analyses used repeated-measures ANOVA, paired t-tests, and linear regression in SPSS. Fluorescence intensities were normalized and binding potential was calculated using a compartment model.
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