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Overview of Flow Cytometry Instrumentation
Publication Name:
Current Protocols in Cytometry
Unit Number:
Unit 1.1
DOI:
10.1002/0471142956.cy0101s39
Online Posting Date:
January, 2007
Abstract
This unit describes the technology in general, to give a feel for the interplay between the various parts of a flow cytometer. Topics include cell preparation, detectors, analysis, and sorting. A brief chronology of flow instrumentation development illustrates the long history of the field.
Figures
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Figure 1.1.1 Schematic diagram of a complete flow cytometer system. -
Figure 1.1.2 Longitudinal cross-sectional view of the flow chamber of a flow cytometer. The sample stream is surrounded by the sheath fluid which confines the cells (black dots) to the center of the chamber. The laser beam is focused onto the cell stream. -
Figure 1.1.3 Arrangement for a simple flow cytometer, containing a single fluorescence detector (photomultiplier) and a photodiode for detecting laser light scattered by a cell. -
Figure 1.1.4 Arrangement for a flow cytometer with dual fluorescence detectors and a scatter detector. Light from two fluorescent probes is separated by the dichroic mirror and optical filters. With the appropriate filters, photomultiplier P1 can also be used to measure light scattered at 90° to the laser beam. -
Figure 1.1.5 Flow cytometer with two excitation beams (lasers) that are separated vertically by 200 µm. The sample stream is imaged at the half mirror, which is used to direct fluorescent light from each beam interaction to a different pair of photomultipliers, each of which has a beam splitter and filter arrangement as in Figure -
Figure 1.1.6 Diagram illustrating the principle of cell sorting. Cells flowing through the system are represented by small black dots. As cells to be sorted approach the end of the solid stream, a charge is applied to the stream. As the drop carrying the cell separates from the stream, the drop carries the charge. Passing between the high-voltage plates, charged drops containing desired cells are deflected into separate collection beakers. Deflection can be left or right, allowing for the simultaneous sorting of two classes of cells. In this illustration, two drops are sorted for each cell.
Videos
Literature Cited
| Literature Cited | |
| Melamed, M.R., Lindmo, T., and Mendelsohn, M.L. (eds.) 1990. Flow Cytometry and Cell Sorting, 2nd ed. Wiley-Liss, New York. | |
| Shapiro, H.M. 2003. Practical Flow Cytometry, 4th ed. John Wiley & Sons, Hoboken, N.J. | |
| Key References | |
| Melamed, M.R., Mullaney, P.F., and Mendelsohn, M.L. (eds.) 1979. Flow Cytometry and Cell Sorting, 1st ed. John Wiley & Sons New York. | |
|
First overall summary of the field, with many authors describing the state of the art as of 1979. Covers applications of the technology as well as instrumentation. | |
|
Melamed, Lindmo, and
Mendelsohn (eds.)
1990.
See | |
|
Second edition with all-new papers, producing true update of the first edition. Also contains extensive history of the field. | |
|
Shapiro, H.M.
2003.
See | |
|
Comprises user's reference manual for the laboratory. Also includes extensive list of current literature and list of key suppliers of instruments, parts, and reagents. | |
| Van Dilla, M.A., Dean, P.N., Laerum, O.D., and Melamed, M.R. 1985. Flow Cytometry: Instrumentation and Data Analysis. Academic Press, London. | |
|
Contains papers by leading experts in the field on both subjects. | |
| Watson, J.U. 2004. Introduction to Flow Cytometry. Cambridge University Press, Cambridge. | |
|
Includes detailed discussion of all aspects of flow cytometry instrumentation and measurement phyics. | |
| Internet Resources | |
| http://www.isac-net.org | |
|
Homepage of the International Society for Analytical Cytology (ISAC). | |
| http://www.cyto.purdue.edu | |
|
Comprehensive cytometry resource. | |
