Lymphocytes - Thymocytes
Immunophenotyping of human and murine lymphocytes as well as murine thymocytes is a common application of multi-colour or polychromatic flow cytometry.
Murine bone marrow preparations can be immuno-phenotyped to facilitate the detection of rare events like stem cells by use of a negative selection cocktail, Sca-1 and c-kit immunophenotyping.
Cell lines at ICMS are commonly immunopheno-typed to study stem cells and apoptosis as well as changes in cell cycle. Megakarkocyte differentiation markers CD41 and CD61 were monitored in the Megakaryocyte cell line CMK in response to retinoic acid. Viability of cells lines is also a common application in flow cytometry using DNA binding fluorescent dyes such as PI and DAPI.
Platelets express the glycoproteins GpIIb/IIIa or CD41-CD61, the fibrinogen receptor, and GpIb or CD42 (von Willibrand Factor receptor) and are commonly used to identify and quantitate platelets. Deficiencies of these glycoproteins result in the respective clinical syndromes Glanzmann's Thrombasthenia and Bernard-Soulier syndrome.
A novel method of detecting antigen specific T cells was developed by producing microsomes from the endoplasmic reticulum (ER) isolated from cells producing the required GFP labelled MHC Class I antigens . These ER derived peptide specific microsomes can then be loaded with specific peptides and used identify antigen specific CD8 T cells.