Department of Dermatology and Venereology, I.M. Sechenov First Moscow State Medical University, Moscow, Russian Federation.
Methods Mol Biol. 2020;2163:109-124. doi: 10.1007/978-1-0716-0696-4_9.
The absolute basophil count (cells/L) can be determined by manual counting of peripheral blood smears or using cell counting chambers as well as by automated hematology analyzers and fluorescence flow cytometry. Manual basophil counting of peripheral blood smears is currently regarded as the reference method, although the limitations of this method (distribution, observer, and statistical errors) are widely recognized. Automated hematology analyzers offer an advantage of larger numbers of counted cells and high throughput but are characterized by inconsistent analytical performance for basophil enumeration. Flow cytometric enumeration of circulating basophils using panels of monoclonal antibodies is being developed as novel candidate reference method for the absolute basophil count in peripheral blood. Basophil counting using fluorescence flow cytometry is characterized by high precision and statistical superiority. Emerging innovative technologies for absolute cell counts include imaging flow cytometry, mass cytometry, and on-chip blood counting, but their analytical performance for absolute basophil counts is yet to be established. Here, we describe various techniques for absolute basophil counting in peripheral blood including manual basophil counts in smears and hemocytometers and flow cytometric methodologies using double-platform, bead-based, and volumetric approaches.
绝对嗜碱性粒细胞计数（细胞/L）可通过手动计数外周血涂片或使用细胞计数室以及自动化血液学分析仪和荧光流式细胞术来确定。目前，手动计数外周血涂片被认为是参考方法，尽管这种方法存在局限性（分布、观察者和统计误差）已被广泛认识。自动化血液学分析仪具有计数更多细胞和高通量的优势，但在嗜碱性粒细胞计数方面的分析性能不一致。使用单克隆抗体面板对循环嗜碱性粒细胞进行流式细胞术计数，正在作为外周血绝对嗜碱性粒细胞计数的新型候选参考方法得到发展。使用荧光流式细胞术计数嗜碱性粒细胞具有高精度和统计学优势。用于绝对细胞计数的新兴创新技术包括成像流式细胞术、质谱流式细胞术和芯片上血液计数，但它们在绝对嗜碱性粒细胞计数方面的分析性能尚待建立。在这里，我们描述了外周血中绝对嗜碱性粒细胞计数的各种技术，包括涂片和血球计数板上的嗜碱性粒细胞手动计数，以及使用双平台、基于珠粒和体积的流式细胞术方法。
