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Solid phase cytometry for detection of rare events

✍ Scribed by Karine Mignon-Godefroy; Jean-Gérard Guillet; Cécile Butor

Publisher
John Wiley and Sons
Year
1997
Tongue
English
Weight
241 KB
Volume
27
Category
Article
ISSN
0196-4763

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✦ Synopsis

We describe a simple and rapid laser scanning device for solid phase cytometry. This system can detect and count fluorescent cells over a 22 mm diameter surface (membrane or glass circle) as well as quantify the fluorescence that they emit. A comprehensive discrimination package includes optical and software parameters and accurately distinguishes between valid signals (e.g., labelled cells) and nonspecific signals (e.g., auto-fluorescent particles and debris). Any event detected may also be easily confirmed by visual observation after transfer of the sample to an epifluorescence microscope fitted with a motorized stage driven by the system.

We show a linear relationship between the amount of fluorescein coupled to the cells and the fluorescence signal of the cells detected. This approach is not destructive and further characterization of the sample may be carried out. We have been able to detect rare cellular events at a frequency of 10 27 in 3 min. Potential applications include monitoring of residual disease in oncology and detection of virusinfected cells circulating at very low frequencies.

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