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Gently Down The Stream: Flow Cytometry As Microscopy

Published online by Cambridge University Press:  02 July 2020

Howard M. Shapiro
Affiliation:
283 Highland Avenue, West Newton, Massachusetts , 02465-2513, USA
Howard M. Shapiro
Affiliation:
283 Highland Avenue, West Newton, Massachusetts , 02465-2513, USA
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Abstract

Flow cytometry is an analytical technique in which optical measurements are made of cells or other biological particles as the cells or particles flow, ideally in single file, in a fluid stream past one or more optical measurement stations. Modern optical flow cytometers typically measure light scattered at small (1-5°) and large (15°-135°) angles to an illuminating laser beam, and fluorescence emitted in three or more discrete spectral bands; the most complex instruments employ three or four spatially separated illuminating beams at different wavelengths and can measure twelve fluorescence signals from each cell or particle analyzed.

Flow cytometry was developed in the 1960's to speed up and automate microspectrophotometry and microfluorometry, which had previously been microscope-based, in order to facilitate development of observer-independent clinical tests which could replace the Papanicolaou smear and the differential leukocyte count. At that time, high-resolution scanning was an extremely slow process; high-intensity sources were restricted to arc lamps, and memory in which images could be stored was expensive.

Type
Correlative Fluorescent Microscopy and Flow Cytometry Techniques (Organized by R. Smith)
Copyright
Copyright © Microscopy Society of America 2001

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References

1.)Shapiro, HM: Practical Flow Cytometry, 3rd Edition. New York, Wiley-Liss, 1995.Google Scholar
2.)Durack, J, Robinson, JP, eds., Emerging Tools for Single Cell Analysis: Advances in Optical Measurement Technologies, New York, Wiley-Liss, 2000.CrossRefGoogle Scholar
3.)Diamond, RA, DeMaggio, S, eds., in Living Color: Protocols in Flow Cytometry and Cell Sorting, Berlin, Springer, 2000Google Scholar