a Centre for Haemophilia and Thrombosis, Department of Clinical Biochemistry , Aarhus University Hospital , Aarhus , Denmark.
Platelets. 2018 Mar;29(2):199-202. doi: 10.1080/09537104.2017.1353684. Epub 2017 Sep 29.
Flow cytometry is an increasingly used method for platelet function analysis because it has some important advantages compared with other platelet function tests. Flow cytometric platelet function analyses only require a small sample volume (3.5 mL); however, to expand the field of applications, e.g., for platelet function analysis in children, even smaller volumes are needed. Platelets are easily activated, and the size of the needle for blood sampling might be of importance for the pre-activation of the platelets. Moreover, to use flow cytometry for investigation of platelet function in clinical practice, a reference interval is warranted. The aims of this work were 1) to determine if small volumes of whole blood can be used without influencing the results, 2) to examine the pre-activation of platelets with respect to needle size, and 3) to establish reference intervals for flow cytometric platelet function assays. To examine the influence of sample volume, blood was collected from 20 healthy individuals in 1.0 mL, 1.8 mL, and 3.5 mL tubes. To examine the influence of the needle size on pre-activation, blood was drawn from another 13 healthy individuals with both a 19- and 21-gauge needle. For the reference interval study, 78 healthy adults were included. The flow cytometric analyses were performed on a NAVIOS flow cytometer (Beckman Coulter, Miami, Florida) investigating the following activation-dependent markers on the platelet surface; bound-fibrinogen, CD63, and P-selectin (CD62p) after activation with arachidonic acid, ristocetin, adenosine diphosphate, thrombin-receptor-activating-peptide, and collagen. The study showed that a blood volume as low as 1.0 mL can be used for platelet function analysis by flow cytometry and that both a 19- and 21-gauge needle can be used for blood sampling. In addition, reference intervals for platelet function analyses by flow cytometry were established.
流式细胞术是一种越来越常用的血小板功能分析方法，因为与其他血小板功能测试相比，它具有一些重要的优势。流式细胞术血小板功能分析仅需要小样本量（3.5 毫升）；然而，为了扩大应用范围，例如，用于儿童的血小板功能分析，甚至需要更小的体积。血小板很容易被激活，并且采血针的大小可能对血小板的预激活很重要。此外，为了在临床实践中使用流式细胞术研究血小板功能，需要建立参考区间。本研究的目的是：1）确定小体积全血是否可以使用而不影响结果，2）检查针的大小对血小板预激活的影响，3）建立流式细胞术血小板功能检测的参考区间。为了检查样本量的影响，从 20 名健康个体中采集了 1.0 毫升、1.8 毫升和 3.5 毫升管中的血液。为了检查针的大小对预激活的影响，从另外 13 名健康个体中用 19 号和 21 号针抽取了血液。为了进行参考区间研究，纳入了 78 名健康成年人。在贝克曼库尔特公司（Beckman Coulter，佛罗里达州迈阿密）的 NAVIOS 流式细胞仪上进行了流式细胞术分析，检测了血小板表面以下激活依赖性标志物：花生四烯酸、瑞斯托霉素、二磷酸腺苷、凝血酶受体激活肽和胶原激活后结合的纤维蛋白原、CD63 和 P-选择素（CD62p）。研究表明，低至 1.0 毫升的血液量即可用于流式细胞术的血小板功能分析，并且可以使用 19 号和 21 号针进行采血。此外，还建立了流式细胞术血小板功能分析的参考区间。
