Methods for Isolation of Tumor-Associated Endothelial Cells for Surface Protein Analysis and Sorting by Flowcytometry

Polychromatic flowcytometry is increasingly used for simultaneously analyzing multiple intracellular and cell-surface proteins on a given cell population. Here we describe a flowcytometry-based method to analyze various proteins on the surface of endothelial cells (which comprise of less than 0.5% of the tumor microenvironment) and concurrently sort the live endothelial cells for the downstream applications such as gene expression by conventional quantitative PCR or by single-cell RNA sequencing.