Development Of Flow Cytometric Technologies For Immune Assays

Roederer, Mario

Abstract

Flow Cytometry is the primary tool in most immunology laboratories for the detailed characterization of lymphocytes and lymphocyte function. The ability to resolve highly complex populations of cells, interrogate the cells one-by-one, make multiple measurements on each cell, and then separate the cells viably for further study make this technology unique and indispensable. Our laboratory has long been involved in pushing this technology forward; we have perhaps the most sophisticated flow cytometer in the world, currently capable of making 20 different measurements on each cell. This platform has proven invaluable in dissecting immune responses to vaccines and viruses. In addition, we are developing novel algorithms for the automated analysis of the highly complex data that results from these experiments.

Funding Agency

• Agency: National Institute of Health (NIH)
• Institute: National Institute of Allergy and Infectious Diseases (NIAID)
• Type: Intramural Research (Z01)
• Project #: 1Z01AI005020-07
• Application #: 7732743
• Support Year: 7
• Fiscal Year: 2008
• Total Cost: $1,080,357

Institution

• Name: National Institute of Allergy and Infectious Diseases
• Country: United States

Publications

Precopio, Melissa L; Butterfield, Tiffany R; Casazza, Joseph P et al. (2008) Optimizing peptide matrices for identifying T-cell antigens. Cytometry A 73:1071-8

Roederer, Mario; Moody, M Anthony (2008) Polychromatic plots: graphical display of multidimensional data. Cytometry A 73:868-74

Mahnke, Yolanda D; Roederer, Mario (2007) Optimizing a multicolor immunophenotyping assay. Clin Lab Med 27:469-85, v

Liu, Jie; Roederer, Mario (2007) Differential susceptibility of leukocyte subsets to cytotoxic T cell killing: implications for HIV immunopathogenesis. Cytometry A 71:94-104

Chattopadhyay, Pratip K; Yu, Joanne; Roederer, Mario (2007) Application of quantum dots to multicolor flow cytometry. Methods Mol Biol 374:175-84

Perfetto, Stephen P; Roederer, Mario (2007) Increased immunofluorescence sensitivity using 532 nm laser excitation. Cytometry A 71:73-9

Chattopadhyay, Pratip K; Price, David A; Harper, Theresa F et al. (2006) Quantum dot semiconductor nanocrystals for immunophenotyping by polychromatic flow cytometry. Nat Med 12:972-7

Parks, David R; Roederer, Mario; Moore, Wayne A (2006) A new ""Logicle"" display method avoids deceptive effects of logarithmic scaling for low signals and compensated data. Cytometry A 69:541-51

Lamoreaux, Laurie; Roederer, Mario; Koup, Richard (2006) Intracellular cytokine optimization and standard operating procedure. Nat Protoc 1:1507-16

Perfetto, Stephen P; Chattopadhyay, Pratip K; Lamoreaux, Laurie et al. (2006) Amine reactive dyes: an effective tool to discriminate live and dead cells in polychromatic flow cytometry. J Immunol Methods 313:199-208