Measurement of nuclear translocation in primary T cells using correlation analysis of images obtained on the ImageStream®

Classical biochemical techniques are semi-quantitative in nature, and do not provide per-cell translocation measurements. Per-cell scoring can be achieved using direct visual observation with a fluorescence microscope, but the variation in extent of nuclear translocation presents a significant challenge to objective image-based measurement, and because manual image acquisition and
quantitative image analysis are time consuming processes, such analyses of nuclear translocation have remained largely qualitative.

We have recently shown that nuclear translocation events within non-adherent tumor cells can be quantified by correlating transcription factor and nuclear images collected in flow using the ImageStream imaging flow cytometer. This application notes details extension of the quantitative technique to the study of nuclear localization within primary human or murine T cells