Dumont L J, Dumont D F
Research and Development, COBE BCT, Inc., Lakewood, Colorado 80215, USA.
Cytometry. 1996 Dec 15;26(4):311-6. doi: 10.1002/(SICI)1097-0320(19961215)26:4<311::AID-CYTO12>3.0.CO;2-U.
The increased demand for leukocyte-reduced platelet products has prompted actions both by blood suppliers and device manufacturers to develop effective quality assurance methods and improved devices for leukocyte-reduced platelet preparation. White cell (WBC) counting methods capable of counting WBCs substantially below 1,000 WBC/mL enhance these activities. The purpose of this study was to enhance and validate a method capable of counting 5-10 WBC/mL in platelets.
We added marker chicken red blood cells (cRBC) at a known concentration to an aliquot of platelet concentrate containing propidium iodide stain solution (PI). A modified BD FACScan was used to analyze this mixture. The ratio of cRBC and WBC events observed was used with the cRBC concentration to calculate the WBC concentration. Validation was performed by analyzing standard curves prepared with leukocyte-depleted platelet diluent. Methods of preparing leukocyte-reduced platelets by apheresis and filtration were evaluated using this method.
The method was linear from 5,000 WBC/mL to 10 WBC/mL (r2 = 0.983). Residual WBCs in the platelet diluent hampered validation below 10 WBC/mL. Repeatability (CV) was 44% at 8 WBC/mL and 6.5% at 1,000 WBC/mL. The FACScan required frequent mixing of the sample and flushing with cleaning agents during acquisition.
This method has good linearity and reproducibility from 10-5,000 WBC/mL for platelets. It is suited for research and development work, and may be a useful adjunct method for quality assurance in the blood center.
对白细胞去除血小板制品需求的增加促使血液供应商和设备制造商采取行动，以开发有效的质量保证方法和改进的白细胞去除血小板制备设备。能够对白细胞计数显著低于1000个白细胞/毫升的白细胞（WBC）计数方法可加强这些工作。本研究的目的是改进并验证一种能够对血小板中5 - 10个白细胞/毫升进行计数的方法。
我们将已知浓度的标记鸡红细胞（cRBC）添加到含有碘化丙啶染色溶液（PI）的血小板浓缩液等分试样中。使用改良的BD FACScan分析仪分析该混合物。观察到的cRBC与WBC事件的比例与cRBC浓度一起用于计算WBC浓度。通过分析用白细胞去除血小板稀释剂制备的标准曲线进行验证。使用该方法评估了通过单采和过滤制备白细胞去除血小板的方法。
该方法在5000个白细胞/毫升至10个白细胞/毫升范围内呈线性（r2 = 0.983）。血小板稀释剂中的残留白细胞阻碍了低于10个白细胞/毫升时的验证。在8个白细胞/毫升时重复性（CV）为44%，在1000个白细胞/毫升时为6.5%。在采集过程中，FACScan分析仪需要频繁混合样品并用清洁剂冲洗。
该方法在10 - 5000个白细胞/毫升范围内对血小板具有良好的线性和重现性。它适用于研发工作，可能是血液中心质量保证的一种有用辅助方法。
