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.

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
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