Using imaging flow cytometry has been possible to optimize an innovative method for studying co-localization of probes by combining the automatic and rapid collection of images with the objective quantification of probe location and intensity.The correlation analysis built in to the IDEAS image analysis software provides robust statistically significant results on large numbers of cells so that phenotypically distinct subsets of cells can be analyzed. In this experiment we use the capabilities of the ImageStreamX system, to measure the transit of a fluorescently labeled antibody-drug conjugate (ADC) through the cellular endocytic pathway. The high spatial resolution afforded by the ImageStreamX system allowed accurate measurement of ADC co-localization to endosomes and lysosomes.
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