Flow Cytometry

SNARF

SNARF is utilized for intracellular pH measurements, and emits differently at different pHs. To label cells, used as an example to show two flow cytometry (to give an idea of double staining - 2 flow graph). Can assess any decreased function by comparing SNARF (dye stain everything) to other dyes and comparing the level of lymphocytes migrating to other compartments, like the lymph. SNARF labeled cells in vitro. It can be used to label cells in vivo. Cell trackers could be used to assess proliferative properties, through cell tracking studies using intravital microscopy (IVM). IVM, through visualization through green fluorescent protein GFP expression on a CD45 e.g., promoter (transgenic), would allow us to see all cells with this receptor expressed, and we could compare levels between before and after antigen stimulation experiments, through counting lymphocytes and extrapolating.

References

1. Janeway, C.A., Travers, P., Walport, M., and Capra, J.D. 1999. Immunobiology: The immune system in health and disease. Garland Publishing, 4th ed., New York, USA

2. Delves, P. and Roitt, I. 1999. Encyclopedia of Immunology. Academic Press Inc., 2nd ed., San Diego, USA

3. 1994. Current Protocols in Molecular Biology. Volume 2. John Wiley & Sons Inc., USA

4. Cruse, J. and Lewis R. 1995. Illustrated Dictionary of Immunology. CRC Press Inc., USA

5. http://biology.berkeley.edu/crl/flowcytometry.shtml

6. Lodish, H., Baltimore, D., Berk, A., Zipursky, Matsudaira, and Darnell, J. 2000. Molecular Cell Biology. American Scientific Books, USA

7. www.spectracell.com/jana.html

8. Andrade, W., Johnston, M. and Hay, J.B. 1998. The relationship of blood lymphocytes to the recirclating lymphocyte pool. Blood 91: 1653-1661.