Count of residual leukocyte numbers in leukocyte depleted products. A sample concentration method for flow cytometry.:A SAMPLE CONCENTRATION METHOD FOR FLOW CYTOMETRY
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The number of residual leukocytes in blood filtered through a leukocyte removal filter was determined by flow cytometric technique after concentrating specimens (a sample concentration method). In this method, propidium iodide (PI) was used to stain the nuclei of leukocytes remaining in the blood. To determine the number of leukocytes in red cell concentrates (RCC), seven times as much PI solution (3500μl) as volume of specimen (500μl) was added, and the suspension was centrifuged. After removal of the supernatant, it was resuspended in 400μl of PI solution. With platelet concentrates (PC), 1.5 times as much PI solution (3000μl) as the volume of specimens (2000μl) was added, and the specimen was centrifuged, the supernatant removed, and the sample resuspended in 400μl of PI solution. The regression line obtained after concentration was: y=0.770x-27 (r=0.986, n=21) for RCC and y=0.755x-20 (r=0.970, n=38) for PC. The mean leukocyte removal rate revealed 99.97±0.02% by filtration of 100ml RCC using a Pall RC 100 filter, and 99.90±0.07% by filtration of 20 units pooled PC using a Pall PL100 filter. The concentration method used in the present study caused no silting in the flow cytometer which can result from insoluble aggregates generated by concentration. It increased the sensitivity for determination of extremely low numbers of leukocytes. This method will also provide an effective means of increasing the sensitivity of other types of flow cytometries.
- 一般社団法人 日本輸血・細胞治療学会の論文
一般社団法人 日本輸血・細胞治療学会 | 論文
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