Immunology

Cells of the innate and adaptive immune system work in concert to protect the body from pathogenic attack. The many different types of immune cells can be distinguished from one another not only by cell surface markers but also by their diverse cellular responses to different pathogens or stimuli. By combining quantitative image analysis tools with the large sample sizes common to flow cytometry, the ImageStream uniquely enables multiple applications within the field of Immunology, including measurement of nuclear translocation of transcription factors, T:APC interactions and accumulation of proteins at the immune synapse, chemokine-induced shape change, and phagocytosis, just to name a few.

Localization of ADAP to the immunological synapse

Ovalbumin-specific accumulation of ADAP at the immune synapse is measured. Without specific peptide ADAP (green) is distributed around the T cell and in the presence of specific peptide ADAP moves to the synapse between the APC and T cell. Quantitation of conjugate formation and representative cells images show that without specific peptide ADAP (green) is distributed around the T cell and in the presence of specific peptide ADAP moves to the synapse between the APC and T cell. See the application note for more details.

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.

Phagocytosis by murine macrophages

Internalization of zymosan (green) by murine RAW cells (orange) identified by immunophenotyping. Phagocytosis is measured as the percentage of cells with internalized zymosan at 15, 30 and 60 minutes at 37 degrees C.

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.