Voevodsky Institute of Chemical Kinetics and Combustion, Novosibirsk, Russian Federation.
State Research Institute of Circulation Pathology, Novosibirsk, Russian Federation.
Anal Methods. 2021 Jul 29;13(29):3233-3241. doi: 10.1039/d1ay00431j.
Analysis of blood platelets encounters a number of different preanalytical issues, which greatly decrease the reliability and accuracy of routine clinical analysis. Modern hematology analyzers determine only four parameters relating to platelets. Platelet shape and dose-dependent activation parameters are outside the scope of commercial instruments. We used the original scanning flow cytometer for measurement of angle-resolved light scattering and the discrete dipole approximation for simulation of light scattering from a platelet optical model, as an oblate spheroid, and global optimization with two algorithms: the DATABASE algorithm to retrieve platelet characteristics from light scattering and the DIRECT algorithm to retrieve dose-dependent activation parameters. We developed the original sampling protocol to decrease spontaneous platelet activation. The new protocol allows us to keep most of the platelets in resting and partially activated states before analysis. The analysis delivers 13 content and morphological parameters of the platelets. To analyze platelet shape change during ADP activation we developed a phenomenological model. This model was applied to the analysis of ADP activation of platelets to give 8 dose-dependent activation parameters. To demonstrate the applicability of the developed protocol and analytical method, we analyzed platelets from five donors. This novel approach to the analysis of platelets allows the determination of 21 parameters relating to their content, morphology and dose-dependent activation.
血小板分析会遇到许多不同的预分析问题，这极大地降低了常规临床分析的可靠性和准确性。现代血液学分析仪仅能确定与血小板相关的四个参数。血小板的形状和剂量依赖性激活参数不在商业仪器的范围内。我们使用原始的扫描流式细胞仪来测量角度分辨光散射，并使用离散偶极近似来模拟血小板光学模型的光散射，将其视为扁球体，并使用两种算法进行全局优化：DATABASE 算法用于从光散射中检索血小板特征，DIRECT 算法用于检索剂量依赖性激活参数。我们开发了原始的采样方案来减少血小板的自发激活。新方案允许我们在分析前将大部分血小板保持在静止和部分激活状态。分析提供了血小板的 13 个含量和形态参数。为了分析 ADP 激活过程中血小板的形状变化，我们开发了一个唯象模型。该模型应用于 ADP 激活血小板的分析，得出了 8 个剂量依赖性激活参数。为了演示所开发方案和分析方法的适用性，我们分析了来自五个供体的血小板。这种分析血小板的新方法可以确定与它们的含量、形态和剂量依赖性激活相关的 21 个参数。
