Flow Cytometry and Cell Sorting

Flow cytometric analysis of molecular, biochemical, genetic and developmental parameters using cellular fluorescence techniques as well as fluorescence-activated (FACS) or magnetic (MACS) cell sorting technologies provide unique options for molecular and cellular biology. In recent years, these technologies have been considerably advanced.
In this second edition, all chapters have been updated according to the recent improvements and modifications. Further, new protocols have been added, such as on magnetic selection, magnetofluorescent liposomes, the cytometry of secreted products and microbead assays, as well as reporter gene assays for cytometry and cell selection. The recent technical developments allow diagnostic differentiation of cells according to specific gene expression, identification of rare disease-associated cells and isolation of well-defined cells at high purity for cell therapies.

Easy-to-follow step-by-step instructions for immediate use at the lab bench protocols with a uniform clear structure and layout, which allow the techniques to be successfully reproduced All necessary practical details and notes as well as a troubleshooting section are included Up-to-date methods written by experts in their fields Includes supplementary material: sn.pub/extras

Contenu
I Flow Cytometry.- 1 Operation of a Flow Cytometer.- II Immunofluorescence.- 2 Conjugation of Fluorochromes, Haptens, and Phycobiliproteins to Antibodies.- 3 Immunofluorescence: Basic Considerations.- 4 Multicolor Immunofluorescence Analysis.- 5 Combined Intracellular and Surface Staining: Immunofluorescence of Cytokines in T Cells.- 6 Cytometry and Sorting of Live Cells According to Their Secreted Products.- 7 Cytometry of Rare Surface Molecules by Magnetofluorescent Liposomes.- 8 Scatchard Analysis by Flow Cytometry.- 9 Microbead Assay for Quantification of Neuronal Adhesion Molecule Interaction by Flow Cytometry.- III DNA and Proliferation.- 10 Preparation and Staining of Fixed Cells for High-Resolution DNA Analysis.- 11 Simultaneous Flow Cytometric Detection of Bromodeoxyuridine Incorporation and Cell Surface Marker Expression.- 12 High-Resolution Cell Cycle Analysis: The Flow Cytometric Bromodeoxyuridine-Hoechst Quenching Technique.- IV Cellular Activation and Biochemistry.- 13 Cell Activation: Indo-1 Ratiometric Analysis of Intracellular Ionized Calcium.- 14 Detection and Purification of Rare Responders by Fixed-Time Flow Cytometry.- 15 Biochemical Parameters of Cells Function.- 16 The FACS-Gal Assay.- 17 Ligand Acidification by Nonadherent Cells.- V Cell Sorting.- 18 Powerful Preselection.- 19 Magnetic Selection of Transfected Cells.- 20 High Gradient Magnetic Cells Sorting.- 21 Setup of a Flow Sorter.- 22 Sorting of Rare Cells.- 23 Fluorescence-Activated Chromosome Sorting.- 24 Analysis and Sorting of Plant Chromosomes by Flow Cytometry.- 25 Large Particle Sorting.- VI Safety.- 26 Biological and Laser Safety.- Appendices: Birgit Mechtold and Andreas Radbruch.- Suppliers.- Cytometry on the Internet.