Quantification of solution-free red blood cell staining by sorption kinetics of Romanowsky stains to agarose gels
Quantification of solution-free red blood cell staining by sorption kinetics of Romanowsky stains to agarose gels
复制标题
通过罗曼诺夫斯基染色剂对琼脂糖凝胶的吸附动力学对无溶液红细胞染色进行定量
DOI：
10.1039/d3ay01431b
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发表时间：
2023
影响因子：
3.1
通讯作者：
Jung, Jangwook P.
中科院分区：
文献类型：
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作者：
Bae, Chae Yun;Esmaeili, Hamid;Zamin, Syed A.;Seol, Min Jeong;Hwang, Eunmi;Beak, Suk Kyung;Song, Younghoon;Bharti, Bhuvnesh;Jung, Jangwook P.
染色红细胞（rbc）的成像和定量对于血液学中识别红细胞和细胞病理学中诊断病变红细胞或寄生虫具有重要意义。罗曼诺夫斯基染色传统上使用阴离子伊红Y和阳离子亚甲基蓝和天蓝色b的混合物在血细胞中产生色调。虽然罗曼诺夫斯基染色已广泛用于细胞病理学，但由于甲醇过早沉淀或蒸发，最终用户在染色中遇到了不同结果的问题，导致基于溶液的罗曼诺夫斯基染色固有的不一致性。在此，我们证明了血液涂片的染色和染色是由琼脂糖凝胶印章的接触时间控制的。虽然染色和染色的程度可以通过显微照片中印记红细胞的色调值来辨别，但琼脂糖凝胶印记吸附和解吸的罗曼诺夫斯基染料分子（特别是伊红Y、亚甲基蓝和蔚蓝B）的定量需要一个可以解释吸附过程的模型。我们从吸附的准二级动力学模型和解吸的一相衰变模型中发现了可预测的罗曼诺夫斯基染料分子的吸附。因此，这里展示的琼脂糖凝胶印花方法可以替代基于溶液的罗曼诺夫斯基染色，具有可预测的吸附量和接触时间。
The imaging and quantification of stained red blood cells (RBCs) are important for identifying RBCs in hematology and for diagnosing diseased RBCs or parasites in cytopathology. Romanowsky staining has been used traditionally to produce hues in blood cells using a mixture of anionic eosin Y and cationic methylene blue and azure B. While Romanowsky stains have been widely used in cytopathology, end-users have experienced problems with varying results in staining due to the premature precipitation or evaporation of methanol, leading to the inherent inconsistency of solution-based Romanowsky staining. Herein, we demonstrate that the staining and destaining of blood smears are controllable by the contact time of agarose gel stamps. While the extent of staining and destaining is discernable by the hue values of stamped red blood cells in micrographs, the quantification of adsorbed and desorbed Romanowsky dye molecules (in particular, eosin Y, methylene blue and azure B) from and to the agarose gel stamps needs a model that can explain the sorption process. We found predictable sorption of the Romanowsky dye molecules from the pseudo-second-order kinetic model for adsorption and the one phase decay model for desorption. Thus, the method of agarose gel stamping demonstrated here could be an alternative to solution-based Romanowsky staining with the predictable quantity of sorption and timing of contact.