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Additional info for Guide to Flow Cytometry Methods
The aerosol conditions presented by a flow cytometer's operation, especially during cell sorting, still further heightens the need for precautions. Some flow cytometers have no containment provisions, while others have the means of producing negative pressure (biohazard containment) or positive pressure (for sterile sorting) within the sorting chamber- these must never be confused (see Chap. 7, Sect. 4). Excellent discussions of various means of biohazard containment in a flow cytometry laboratory have been presented by Muirhead et al.
1: Intracellular Glutathione Using Monochlorobimane 175 Page ix References 176 12. 1: Anisotropy Measurement for Membrane Fluidity 187 References 187 13. 1: Three Tube Method for Estimation of Number of Receptors 201 References 202 Recommended Reading 203 14. 1: Membrane Potential Changes 208 References 209 15. 1 Flow Cytometers In 1956, Coulter developed a commercially successful device for counting blood cells by means of differences in electrical conductivity between the cells and their medium (the Model A Coulter Counter).
They are capable of dual laser beam excitation and simultaneous measurement of several different fluorescence wavelengths, forward angle and right angle light scatter, Coulter volume, time as a parameter, and time of flight measurements. They can also physically separate (sort) subpopulations of cells on the basis of any parameter or combination of parameters. In a typical flow cytometer, stained cells in suspension are made to flow in a laminar manner by a suitable hydrodynamic system, such that they line up in single file, like beads on a string.