Abstract

Traditional immunofluorescence (IF) imaging assays are limited to the detection of just a few markers due to spectral overlap of fluorescent emission bands. Furthermore, standard fluorescent imaging instruments have a dynamic range that is too narrow to capture the full range of expression values seen in biology, precluding the accurate quantification of single-cell target expression. Here we describe a protocol for detection and quantification of dozens of protein targets with single-cell quantitative precision using an iterative staining approach called ChipCytometry™.

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