A reversible multi-well chamber for incubation of cultured cells with small volumes: application to screening of hybridoma fusions using immunofluorescence microscopy.
Immunofluorescence microscopy is a powerful technique for detecting the location of surface and intracellular antigens in individual cells. However, using standard methods, processing large numbers of samples for immunofluorescence is cumbersome and difficult. To simplify greatly this process, we have developed a chamber that reversibly creates multiple small wells in a large (150 mm) tissue culture dish. This device allows the rapid and convenient processing of hundreds of samples each of 100 microliters volume. Each sample is examined using a short working distance, high numerical aperture immersion objective for maximum sensitivity and resolution. This apparatus makes immunofluorescence a practical method for the primary screening of hybridoma clones.
Ultrastructural Cytochemistry Section, National Cancer Institute, Bethesda, MD 20892.
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Willingham
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Pastan
D000911:Antibodies, Monoclonal / Q000032:analysis; D000954:Antigens, Surface / Q000032:analysis; D002478:Cells, Cultured / Q000032:analysis*; D005455:Fluorescent Antibody Technique / Q000295:instrumentation*; D006825:Hybridomas / Q000276:immunology; D008856:Microscopy, Fluorescence; D012235:Rhodamines
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D012235:Rhodamines