SpringerImages - Schematic diagram illustrating the principles of flow cytometry and sorting. Fluorescently stained cells are introduced into the flow chamber and forced to flow one by one through an intense light beam that excites the fluorescent dye. The resulting fluorescence is collected and recorded as a measure of the amount of bound dye. Since the dye is usually chosen to bind selectively and stoichiometrically to a specific cellular component (DNA in this example), the recorded fluorescence intensity is used as a measure of the amount of the stained cellular component. Cells can be measured at rates of up to 20,000/s with an accuracy approaching 2% (46). After analysis, the cells flow out of a small orifice in a thin jet. The jet is forced to break into uniform droplets by the action of a vibrating piezoelectric transducer mounted on top of the flow chamber. Cells possessing distinctive features (mid-S-phase DNA content in this example) are selected for sorting by applying a charge to the droplets carrying these cells. The charged droplets are separated from the uncharged droplets during passage through an intense electric field and can be collected for further analysis (30

Caption

Fig 1

Schematic diagram illustrating the principles of flow cytometry and sorting. Fluorescently stained cells are introduced into the flow chamber and forced to flow one by one through an intense light beam that excites the fluorescent dye. The resulting fluorescence is collected and recorded as a measure of the amount of bound dye. Since the dye is usually chosen to bind selectively and stoichiometrically to a specific cellular component (DNA in this example), the recorded fluorescence intensity is used as a measure of the amount of the stained cellular component. Cells can be measured at rates of up to 20,000/s with an accuracy approaching 2% (46). After analysis, the cells flow out of a small orifice in a thin jet. The jet is forced to break into uniform droplets by the action of a vibrating piezoelectric transducer mounted on top of the flow chamber. Cells possessing distinctive features (mid-S-phase DNA content in this example) are selected for sorting by applying a charge to the droplets carrying these cells. The charged droplets are separated from the uncharged droplets during passage through an intense electric field and can be collected for further analysis (30–32,39,55).

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The principles of flow cytometry and sorting are illustrated in Fig.

The insert in Fig.

During sorting, the cells are analyzed flow cytometrically, then ejected into air in a thin liquid jet (see Fig.

In this way, cells selected according to their cytometric properties (e.g., mid-S phase in Fig.

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