Validation of Nanobody and Antibody Based In Vivo Tumor Xenograft NIRF-imaging Experiments in Mice Using Ex Vivo Flow Cytometry and Microscopy

作者： Peter Bannas*1, Alexander Lenz*1,2, Valentin Kunick1,2, William Fumey1,2, Björn Rissiek2, Joanna Schmid1,2, Friedrich Haag2, Axel Leingärtner3, Martin Trepel4, Gerhard Adam1, Friedrich Koch-Nolte2 1Department of Diagnostic and Interventional Radiology,University Medical Center, Hamburg, 2Institute of Immunology,University Medical Center, Hamburg, 3University Cancer Center Hamburg,University Medical Center, Hamburg, 4Department of Oncology and Hematology,University Medical Center, Hamburg

简介：

Overview

This protocol outlines the steps for ex vivo validation of in vivo near-infrared fluorescence xenograft imaging in mice using fluorophore labeled nanobodies and conventional antibodies. The method allows for comprehensive imaging comparisons and helps address key questions in tumor imaging.

Key Study Components

Area of Science

• Neuroscience
• Immunology
• Oncology

Background

• Near-infrared fluorescence imaging is a valuable tool in tumor research.
• Fluorophore labeled antibodies can enhance imaging specificity.
• Ex vivo validation is crucial for confirming in vivo imaging results.
• This method provides a multimodal approach to imaging.

Purpose of Study

• To validate in vivo imaging results using ex vivo techniques.
• To assess the specificity of fluorophore labeled constructs.
• To improve tumor localization and monitoring of therapies.

Methods Used

• Administration of labeled antibodies to mice with xenografts.
• In vivo imaging followed by ex vivo analyses.
• Flow cytometry and fluorescence microscopy for validation.
• Use of specific staining techniques to identify cell types.

Main Results

• Specific labeling of antigen-positive tumor cells was achieved.
• Conventional antibodies showed weaker and non-specific staining.
• Ex vivo analyses confirmed in vivo imaging results.
• The method allows for rapid imaging and analysis within a day.

Conclusions

• This protocol provides a reliable method for validating imaging results.
• Fluorophore labeled constructs enhance specificity in tumor imaging.
• The approach can be adapted for further biodistribution studies.

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