Samples isolated by flow cytometry
In recent years, important technological developments have provided new means to study fundamental aspects in biology and pathology. The emergence of DNA microarray technology enables us to analyze the expression levels of thousands of genes within cells, and to explore the underlying genetic causes of many human diseases. Moreover, the development of global PCR techniques now enables 1 to 10 million fold mRNA amplifications, and therefore microarray analysis from limited biological materials. Fluorescence Activated Cell Sorting (FACS) offers us the opportunity to separate cell populations according to their phenotypic or physical attributes, including rare cell types from heterogeneous mixtures, and to purify these for further characterization. In this chapter we discuss the possibility of combining these major technological advances in order to perform gene expression profiling on isolated cells of interest. The synergy of FACS and microarray technologies is bound to further our capacity to unveil the secrets of life.
KeywordsFluorescence Activate Cell Sort Perform Gene Expression Density Gradient Separation mRNA Amplification Subset Accord
Unable to display preview. Download preview PDF.
- 3.Shapiro, H.M. 2003. Practical Flow Cytometry, 4th edition. Wiley-Liss, HobokenGoogle Scholar
- 4.Perfetto, S.P, P.K. Chattopadhyay, M. Roederer. 2004. Seventeen-colour flow cytometry: unravelling the immune system. Nat Rev Immunol 4: 648–655Google Scholar
- 9.Romero, P, P.R. Dunbar, D. Valmori, M. Pittet, G.S. Ogg, D. Rimoldi, J.L. Chen, D. Lienard, J.C. Cerottini, V. Cerundolo. 1998. Ex vivo staining of metastatic lymph nodes by class I major histocompatibility complex tetramers reveals high numbers of antigen-experienced tumor-specific cytolytic T lymphocytes. J Exp Med 188: 1641–1650PubMedCrossRefGoogle Scholar
- 20.Boutboul, F, D. Puthier, V. Appay, O. Pelle, H. Ait-Mohand, B. Combadiere, G. Carcelain, C. Katlama, S.L. Rowland-Jones, P. Debre et al. 2005. Modulation of interleukin-7 receptor expression characterizes differentiation of CD8 T cells specific for HIV, EBV and CMV. Aids 19: 1981–1986PubMedCrossRefGoogle Scholar
- 26.Appay, V, A. Bosio, S. Lokan, Y. Wiencek, C. Biervert, D. Kusters, D. Devevre, D.E. Speiser, P. Romero, N. Rufer, S. Leyvraz. 2007. Sensitive gene expression profiling of human T-cell subsets reveals parallel post-thymic differentiation for CD4+ and CD8+ lineages. J Immunol; in pressGoogle Scholar