真菌六胺银染色技术的优化

Optimization of the periodic acid-silver metheramine staining for fungi

目的探索真菌六胺银染色的优化方案,寻求染色效果佳且环保简便的染色试剂。方法选取已知真菌感染的阳性病理组织,分别制作成蜡块,通过对比试验,比较不同氧化剂、六胺银染液不同配制方法、不同衬染试剂对六胺银染色质量的影响。结果真菌感染组织切片使用2%高碘酸水溶液氧化,染色步骤较使用三氧化铬溶液氧化简便,且三氧化铬为高毒致癌物,高碘酸毒性较低,故选择2%高碘酸溶液作为氧化剂更优;六胺银改良配制重复利用法配制六胺银液较传统配制法简便,对染液的利用率高,可以重复回收使用,较节约成本;1%亮绿染液与苏木素染液均是教材推荐的衬染剂,但亮绿染液的染色效果显著优于苏木素染液。结论真菌感染组织切片使用2%高碘酸水溶液作为氧化剂、六胺银改良配制重复利用法配制的六胺银液染色、1%亮绿染液衬染为较佳方案。

Objective To explore the optimal scheme of periodic acid-silver metheramine(PASM)for fungi,seeking a staining reagent that provides excellent staining results while being environmentally friendly and simple to use.Methods Positive pathological tissues infected with known fungi were selected and embedded in wax blocks.A comparative test was conducted to evaluate the impact of different oxidants,various preparation methods for the PASM staining solution,and different counterstain reagents on the staining quality.Results Fungal-infected tissue sections were oxidized using a 2%periodate water solution.This staining process was simpler compared to the use of chromium trioxide solution,which is highly toxic and carcinogenic.Periodate was less toxic,the choice of a 2%periodate solution as the oxidant was considered more favorable.The modified formulation and reuse method of the PASM stain simplified the preparation of the staining solution compared to the traditional method,resulting in higher utilization rates and cost savings.While both 1%bright green staining solution and hematoxylin staining solution are recommended counterstains in textbooks,the bright green staining solution exhibited significantly better staining effects than hematoxylin.Conclusion For fungal-infected tissue sections,the optimal staining method involves using a 2%periodic acid aqueous solution as the oxidant,staining with the modified PASM stain prepared using the reuse method,and counterstaining with 1%bright green solution.