Winzek C, Plieninger P, Baumgärtel H
Histochemistry. 1987;86(4):421-6. doi: 10.1007/BF00495004.
A new method to analyze staining processes in single cells of histochemical and cytochemical specimens in situ is described. The combination of a microscope photometer with a perfusion cuvette developed in our laboratory allows the continuous observation of a cell during the staining process. The flow rate dependence of the staining process has been examined demonstrating the strong suppression of the diffusional boundary layer adjacent to the cell surface by sufficiently high flow rates. Experiments to find optimal conditions for the kinetic analysis of the staining reaction of nuclei in lymphocytes, neutrophile granulocytes and monkey kidney cells with thionin are described. Half-staining times of the binding of monomer dye molecules and aggregates to nuclei have been calculated; they depend on the pretreatment of the cells. The addition of electrolytes decreases the rate of staining. The formation of aggregates obeys approximately a first-order reaction law and the binding of monomers provides an order of reaction of n = 0.5.
本文描述了一种原位分析组织化学和细胞化学标本单细胞染色过程的新方法。我们实验室开发的显微镜光度计与灌注比色皿相结合，能够在染色过程中持续观察细胞。研究了染色过程对流速的依赖性，结果表明足够高的流速能强烈抑制细胞表面附近的扩散边界层。本文还介绍了寻找用硫堇对淋巴细胞、中性粒细胞和猴肾细胞核染色反应进行动力学分析的最佳条件的实验。计算了单体染料分子和聚集体与细胞核结合的半染色时间；它们取决于细胞的预处理。添加电解质会降低染色速率。聚集体的形成大致遵循一级反应定律，单体的结合反应级数为n = 0.5。
