Graham J S, Bryant M A, Kirkpatrick L J, Moltrup D L
U.S. Army Medical Research Institute of Chemical Defense, Aberdeen Proving Ground, Maryland 21010-5425.
Biotech Histochem. 1994 May;69(3):121-6. doi: 10.3109/10520299409106273.
Multiple skin sections from three nonhuman primates (Macaca mulatta) and three hairless guinea pigs (Cavia porcellus) were stained with 12 different histologic stains to determine whether mast cells could be selectively stained for morphometric analysis using an image analysis system (IAS). Sections were first evaluated with routine light microscopy for mast cell granule staining and the intensity of background staining. Methylene blue-basic fuchsin and Unna's method for mast cells (polychrome methylene blue with differentiation in glycerin-ether) stained mast cell granules more intensely than background in both species. Toluidine blue-stained sections in the guinea pig yielded similar results. Staining of the nuclei of dermal connective tissue was enhanced with the methylene blue-basic fuchsin and toluidine blue stains. These two stains, along with the Unna's stain, were further evaluated on an IAS with and without various interference filters (400.5-700.5 nm wavelengths). In both the methylene blue-basic fuchsin and toluidine blue stained sections, mast cell granules and other cell nuclei were detected together by the IAS. The use of interference filters with these two stains did not distinguish mast cell granules from stained nuclei. Unna's stain was the best of the 12 stains evaluated because mast cell granule staining was strong and background staining was faint. This contrast was further enhanced by interference filters (500.5-539.5 nm) and allowed morphometric measurements of mast cells to be taken on the IAS without background interference.
对三只猕猴（Macaca mulatta）和三只无毛豚鼠（Cavia porcellus）的多个皮肤切片用12种不同的组织学染色剂进行染色，以确定肥大细胞是否可以使用图像分析系统（IAS）进行选择性染色，用于形态计量分析。首先用常规光学显微镜评估切片的肥大细胞颗粒染色和背景染色强度。亚甲蓝-碱性品红以及Unna氏肥大细胞染色法（多色亚甲蓝在甘油-乙醚中分化）在两种动物中对肥大细胞颗粒的染色都比背景染色更深。豚鼠的甲苯胺蓝染色切片也得到了类似的结果。亚甲蓝-碱性品红和甲苯胺蓝染色增强了真皮结缔组织细胞核的染色。这两种染色剂以及Unna氏染色剂，在有无各种干涉滤光片（波长400.5 - 700.5 nm）的情况下，进一步在IAS上进行评估。在亚甲蓝-碱性品红和甲苯胺蓝染色的切片中，IAS同时检测到了肥大细胞颗粒和其他细胞核。使用干涉滤光片并不能将这两种染色中的肥大细胞颗粒与染色的细胞核区分开来。Unna氏染色剂是所评估的12种染色剂中最好的，因为肥大细胞颗粒染色强而背景染色淡。干涉滤光片（500.5 - 539.5 nm）进一步增强了这种对比度，使得在IAS上可以在无背景干扰的情况下对肥大细胞进行形态计量测量。
