Multiplexing Signal Transduction Measurement

Increasing the number of samples which can be run simultaneously is desirable for high content analysis and screening applications including extensive dose and time course studies. To increase sample throughput, the fluorescent cell barcoding method previously reported by Krutzik & Nolan has been adapted for the ImageStream® to be used in combination with the additional high content analysis and information gained when using imaging flow cytometry over standard flow cytometry. The method involves labeling samples with up to three different Alexa Fluor® succinimidyl ester dyes in different concentrations (i.e., intensities) followed by staining with the markers of interest. In this study nuclear translocation of NF-κB was measured in THP1 cells on a per cell basis in an automated and quantitative manner in up to 64 samples simultaneously.