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Published on: July 6, 2022
Protocol for spatial profiling of immune cells in the mouse cornea and conjunctiva using NanoString GeoMx DSP and
Lois Kim1, Karthikeyan Ramasamy1, Ajay Sharma1
1Department of Biomedical and Pharmaceutical Sciences, Chapman University School of Pharmacy, Chapman University, Irvine, CA 92618, USA.
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Quantification of immune cells and their activation status in a spatially specific manner in a healthy or diseased tissue is a valuable tool in immunology. Here, we present a protocol for the spatial profiling of immune cells in the mouse cornea and conjunctiva. We describe steps for immunostaining with photocleavable oligonucleotide-conjugated antibodies, selecting regions of interest (ROIs), and collecting UV-cleaved oligonucleotides using GeoMx DSP. We then detail procedures for hybridizing collected oligonucleotides, cartridge blotting using nCounter Pro Prep, data acquisition, and data analysis.
