Flow cytometry chamber with 4 pi light collection suitable for epifluorescence microscopes.

A compact, solid, spherico-ellipsoidal chamber (SEC), which has approaching 4 pi ("all around") light collection, has been developed for flow cytometry. This was mounted onto the stage of a standard fluorescence photomicroscope, and the camera was replaced by a photomultiplier. Both components can be added or removed in minutes. The increased light collection efficiency of the SEC (about 85%) compared with about 4% from standard chambers enabled a fluorescence microscope with a 50 W mercury vapour lamp to "double" as a flow cytometer. The system was tested with microbeads and cells stained for DNA with ethidium bromide, and results were comparable to those obtained with our laser-based instrument.