Morikawa Shunichi, Sato Azusa, Ezaki Taichi
Department of Anatomy and Developmental Biology, Tokyo Women's Medical University, 8-1 Kawada-Cho, Shinjuku, Tokyo, Japan.
Department of Chemistry, Tokyo Women's Medical University, 8-1 Kawada-Cho, Shinjuku, Tokyo, Japan.
Microscopy (Oxf). 2018 Dec 1;67(6):331-344. doi: 10.1093/jmicro/dfy037.
Although conventional toluidine blue staining is a common technique used for rapid observation of semithin sections prior to transmission electron microscopy, it is monochromatic and insufficient for accurate identification of different tissue components by light microscopy. Additionally, polychromatic staining methods generally require step-by-step processes involving different dyes, and it is often difficult to balance the color tone of each step. In this study, we developed a simple polychromatic staining method for epoxy-embedded tissue sections. We stained preheated sections with an aqueous ethanol solution of azure B and basic fuchsin, with the addition of sodium tetraborate to enhance the staining efficacy. We optimized various staining conditions to enable sufficient coloration easily and consistently in a single, rapid staining step, using a single staining-mixture solution. Our method enabled clear differentiation of various tissue structures according to color tone and stain intensity, thereby facilitating the detection of fine structural differences, including various organelle and inclusion bodies. This technique represents a simple polychrome-staining method to allow more informative and convincing histological investigation in various fields of research and education.
尽管传统的甲苯胺蓝染色是一种常用于在透射电子显微镜检查之前快速观察半薄切片的技术，但它是单色的，通过光学显微镜难以准确识别不同的组织成分。此外，多色染色方法通常需要涉及不同染料的分步过程，而且往往难以平衡每个步骤的色调。在本研究中，我们开发了一种用于环氧树脂包埋组织切片的简单多色染色方法。我们用含有天青B和碱性品红的乙醇水溶液对预热的切片进行染色，并添加硼砂以提高染色效果。我们优化了各种染色条件，以便在单个快速染色步骤中使用单一染色混合溶液轻松且一致地实现充分染色。我们的方法能够根据色调和染色强度清晰区分各种组织结构，从而便于检测细微的结构差异，包括各种细胞器和包涵体。该技术代表了一种简单的多色染色方法，可在各种研究和教育领域进行更具信息性和说服力的组织学研究。
