Allison Robin W, Velguth Karen E
Department of Veterinary Pathobiology, Center for Veterinary Health Sciences, Oklahoma State University, Stillwater, Oklahoma, USA.
Vet Clin Pathol. 2010 Mar;39(1):99-104. doi: 10.1111/j.1939-165X.2009.00187.x. Epub 2009 Sep 28.
Romanowsky stains are used routinely by veterinary clinical pathology laboratories for cytologic and blood film evaluations. Automated stainers are available for both aqueous and methanolic Romanowsky stains. Mast cell granules and canine distemper virus inclusions are known to stain differently by these 2 methods, but we have noticed differences in the staining characteristics of other granulated cells.
The aim of this study was to investigate and document the variable appearance of basophils and large granular lymphocytes in blood films stained using aqueous and methanolic Romanowsky methods.
Cytologic preparations from 1 canine mast cell tumor and blood films from 8 dogs, 1 cat, 1 rabbit, and 1 ostrich were stained using an automated aqueous stain (Aerospray 7120, with and without a predip fixative) and an automated methanolic stain (Hematek). Staining quality and intensity of the cytoplasmic granules in mast cells, basophils, and large granular lymphocytes was evaluated subjectively.
Cytoplasmic granules of mast cells, basophils, and large granular lymphocytes stained poorly or not at all with the automated aqueous stain but stained prominently and were readily identified with the automated methanolic stain. Use of the predip fixative with the Aerospray method improved the visibility of basophil granules but not mast cell granules, and had a variable affect on the visibility of granules in large granular lymphocytes.
Clinical pathologists should be aware of the staining methodology used on the slides they evaluate to avoid incorrect interpretation of granulated cell populations.
兽医临床病理实验室常规使用罗曼诺夫斯基染色法进行细胞学和血涂片评估。水性和甲醇性罗曼诺夫斯基染色均有自动染色仪可供使用。已知肥大细胞颗粒和犬瘟热病毒包涵体在这两种方法下染色不同，但我们注意到其他颗粒细胞的染色特征也存在差异。
本研究旨在调查并记录使用水性和甲醇性罗曼诺夫斯基方法染色的血涂片中嗜碱性粒细胞和大颗粒淋巴细胞的不同外观。
用自动水性染色仪（Aerospray 7120，有无预浸固定剂）和自动甲醇染色仪（Hematek）对1例犬肥大细胞瘤的细胞制备物以及8只犬、1只猫、1只兔和1只鸵鸟的血涂片进行染色。主观评估肥大细胞、嗜碱性粒细胞和大颗粒淋巴细胞中细胞质颗粒的染色质量和强度。
肥大细胞、嗜碱性粒细胞和大颗粒淋巴细胞的细胞质颗粒在自动水性染色中染色不佳或根本不染色，但在自动甲醇染色中染色显著且易于识别。Aerospray方法使用预浸固定剂可提高嗜碱性粒细胞颗粒的可见性，但不能提高肥大细胞颗粒的可见性，对大颗粒淋巴细胞中颗粒的可见性影响不一。
临床病理学家应了解他们所评估玻片的染色方法，以避免对颗粒细胞群体的错误解读。
