| Budget Amount *help |
¥7,200,000 (Direct Cost: ¥7,200,000)
Fiscal Year 1991: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1990: ¥6,000,000 (Direct Cost: ¥6,000,000)
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| Research Abstract |
Flow cytometry has been widely and extensively used for quantitative cystochmistry in research works and clinical diagnosis. But the procedures for different combination of staining are sometimes laborious and delicate. The aim of the present study is to develop a fullautomated cell workstation for performing various combination of fluorescent staining for flow cytometry. This cell workstation is equipbed with a centrifugal disk carrying ten seats for 5 ml test tubes containing free cell superysion, ten reagent dispensing nozzles which are accessible to the tubes from upward, ten filtration nozzles with mesh filters which are also accessible to the tubes from upward, and three incubation baths of which temperature can be preset independently controlles by heating and cooling sinks which are accessible to the tubes from downward. Ten mixers which are accessible. to the tubes from upward, and washing rooms for the filtration nozzles are also equipped with. All steps of centrifugation, aspiration or pumping out, and filtration of supernatant, reagent dispensing and resuspension, temperature control, mixing solution, and nozzle washing are controlled by a personal computer (PC ; 98019NEC$Japan). The accuracy of the temperature control at 30' C or 370C is kept within + O. S*C for 1 h. The protocols for (1) making free cell suspension from unfixed raw tissue material, (2) quantitative DNA staining with Feulgen reactions DAPI or PI staining procedure, (3) fluorescent acridine orange (AO) staining for differential staining of cancerous cells, and (4) quantitative analysis of Feulgen hydrolysis curve for detecting DNA damage and DNAinstability, were developed and the softwares for automatically performing the protocols were completed.
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