Book contents
- Frontmatter
- Contents
- List of contributors
- List of abbreviations
- 1 Principles of flow cytometry
- 2 Introduction to the general principles of sample preparation
- 3 Fluorescence and fluorochromes
- 4 Quality control in flow cytometry
- 5 Data analysis in flow cytometry
- 6 Laser scanning cytometry: application to the immunophenotyping of hematological malignancies
- 7 Leukocyte immunobiology
- 8 Immunophenotypic analysis of leukocytes in disease
- 9 Analysis and isolation of minor cell populations
- 10 Cell cycle, DNA and DNA ploidy analysis
- 11 Cell viability, necrosis and apoptosis
- 12 Phagocyte biology and function
- 13 Intracellular measures of signalling pathways
- 14 Cell–cell interactions
- 15 Nucleic acids
- 16 Microbial infections
- 17 Leucocyte cell surface antigens
- 18 Recent and future developments: conclusions
- Appendix
- Index
- Plate section
13 - Intracellular measures of signalling pathways
Published online by Cambridge University Press: 06 January 2010
- Frontmatter
- Contents
- List of contributors
- List of abbreviations
- 1 Principles of flow cytometry
- 2 Introduction to the general principles of sample preparation
- 3 Fluorescence and fluorochromes
- 4 Quality control in flow cytometry
- 5 Data analysis in flow cytometry
- 6 Laser scanning cytometry: application to the immunophenotyping of hematological malignancies
- 7 Leukocyte immunobiology
- 8 Immunophenotypic analysis of leukocytes in disease
- 9 Analysis and isolation of minor cell populations
- 10 Cell cycle, DNA and DNA ploidy analysis
- 11 Cell viability, necrosis and apoptosis
- 12 Phagocyte biology and function
- 13 Intracellular measures of signalling pathways
- 14 Cell–cell interactions
- 15 Nucleic acids
- 16 Microbial infections
- 17 Leucocyte cell surface antigens
- 18 Recent and future developments: conclusions
- Appendix
- Index
- Plate section
Summary
Introduction
This chapter reviews and discusses two different flow cytometric approaches to the study of cellular physiology (defined as studies of the dynamic state of a cell population). The first approach uses fixed and permeabilised cells to detect and quantify epitopes on important regulatory molecules as a function of cell cycle phase or stage in the cell death process at a single time point per sample. This approach is static and the time domain, and consequently the physiological state, is inferred. The second approach uses live, dying or dead cells in a native state that have been stained with fluorescent molecules that ‘report’ the physiological state. These measurements are dynamic and fluctuate in real time.
The discussion and examples herein are biased towards the signalling pathways in cancer cells that, in aggregate, define the oncogenic phenotype. Published studies of signalling pathways have increased steadily since the early 1980s, but advances that provide a more complete picture of pathways as interacting nodal networks are more recent. At present, the exact definition of a pathway/network is not very precise. The following is a definition within the context of this chapter: ‘the aggregate biochemical process by which some particular cell behaviour (phenotype) is regulated that includes all chemical changes in the “resting state” that affect the cell phenotype significantly when components of the pathway are modulated.’
Logically, all components within the cell are interconnected to some degree; therefore, modulating one component will probably have some effect on all others. If the regulatory pathways are a multidimensional set of interconnected reactions, one area can only be viewed in isolation by invoking an arbitrary cut-off where the magnitude of an effect on another reaction is deemed insignificant.
- Type
- Chapter
- Information
- Cytometric Analysis of Cell Phenotype and Function , pp. 231 - 246Publisher: Cambridge University PressPrint publication year: 2001