Axelsson L G, Ahlstedt S
Department of Pharmacology, Pharmacia AB, Uppsala, Sweden.
J Immunol Methods. 1990 Mar 9;127(2):271-7. doi: 10.1016/0022-1759(90)90078-a.
Treatment of paraffin or cryostat sections from rat, rabbit and man, with fluorescein isothiocyanate (FITC) in buffer (1 microgram/l), resulted in visualization of highly fluorescent leukocytes with a granular staining in light microscopy. FITC staining of paraffin-embedded colonic tissue was compared with three other staining methods, i.e., (1) FITC-conjugated anti-neutrophil antibodies, (2) endogenous peroxidase with 3,3-diaminobenzidine, and (3) FITC-conjugated Lotus tetragonolobus lectin. Good correlations between FITC staining and the three other methods were found (r:0.97, 0.98 and 0.98; P less than 0.001). Inhibition of receptors for the FITC conjugates prior staining, by preincubation with unconjugated anti-neutrophil antibodies or L-fucose, abolished the staining with antibodies and lectin. This study shows that staining with unlabeled FITC represents a quick and convenient method for the estimation of phagocytic cells (polymorphonuclear cells and macrophages) in histological specimens.
用缓冲液(1微克/升)中的异硫氰酸荧光素(FITC)处理大鼠、兔子和人类的石蜡切片或低温恒温器切片,在光学显微镜下可观察到高度荧光的白细胞呈现颗粒状染色。将石蜡包埋的结肠组织的FITC染色与其他三种染色方法进行比较,即:(1)FITC偶联的抗中性粒细胞抗体,(2)用3,3-二氨基联苯胺检测内源性过氧化物酶,(3)FITC偶联的百脉根凝集素。发现FITC染色与其他三种方法之间具有良好的相关性(r分别为0.97、0.98和0.98;P<0.001)。在染色前,通过与未偶联的抗中性粒细胞抗体或L-岩藻糖预孵育来抑制FITC偶联物的受体,可消除抗体和凝集素的染色。本研究表明,用未标记的FITC染色是一种快速简便的方法,用于评估组织学标本中的吞噬细胞(多形核细胞和巨噬细胞)。