Cell Signaling

Molecular translocation of transcription factors from the cytoplasm to the nucleus is a pivotal event in many processes critical to cellular activation, differentiation, and host defense. Existing methods used to measure nuclear translocation have significant limitations. The ImageStream overcomes these limitations by automatic, rapid collection of images that can be objectively quantified in solution. The ability to numerically score large numbers of automatically acquired images is ideally suited to the analysis of nuclear translocation within many cell types including small, primary immune system cells.

HIV-specific translocation of NFAT

HIV-specific nuclear localization of NFAT was measured in HIV-experienced T cells from peripheral blood of HIV+ patients. HIVgag peptide specifically stimulates NFAT (green) to move to the nucleus (red) in HIV-tetramer positive cells (orange). The Similarity score correlates DRAQ5 nuclear stain with the NFAT signal. The higher the Similarity score, the more translocation is visualized in the example images from each quadrant.

NF-kB translocation in T:APC conjugates

Measurement of NFkB translocation in transgenic T cells which are in contact with antigen presenting cells (APC) and specific peptide. Specific T cell:APC conjugates are identified and translocation of NFkB from the cytoplasm to the nucleus specifically within the T cells is measured in the presence (shown) or absence of peptide. See the application note for more details.