Borzova Elena, Dahinden Clemens A
BMRC, University of East Anglia, Norwich, UK.
Methods Mol Biol. 2014;1192:87-100. doi: 10.1007/978-1-4939-1173-8_7.
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 ImageStream technology 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/升）可以通过对外周血涂片进行手工计数或使用细胞计数板来确定，也可以通过自动血液分析仪和荧光流式细胞术来测定。目前，对外周血涂片进行嗜碱性粒细胞手工计数被视为参考方法，尽管该方法的局限性（分布、观察者和统计误差）已广为人知。自动血液分析仪具有计数细胞数量多和高通量的优势，但在嗜碱性粒细胞计数方面的分析性能不一致。使用单克隆抗体组合通过流式细胞术对循环嗜碱性粒细胞进行计数正在被开发为外周血绝对嗜碱性粒细胞计数的新型候选参考方法。使用荧光流式细胞术进行嗜碱性粒细胞计数具有高精度和统计学优势。新兴的绝对细胞计数创新技术包括ImageStream技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未确定。在此，我们描述了外周血中绝对嗜碱性粒细胞计数的各种技术，包括涂片和血细胞计数器中的手工嗜碱性粒细胞计数以及使用双平台、基于微珠和体积法的流式细胞术方法。
