Shimabukuro T, Yamamoto M, Yoshihiro S, Matsuyama H, Yamamoto N, Sakatoku J
Dept. of Urology, Yamaguchi University School of Medicine.
Gan To Kagaku Ryoho. 1989 Apr;16(4 Pt 1):839-44.
In spite of the development of a staining principle and the method for simultaneous flow cytometric bromodeoxyuridine (BrdU)/DNA analysis, this technic was seldom used clinically, because, it can not be applied for the study of solid tumors. In this paper, we described a new method to overcome this problem by combining proteolytic enzyme digestion with previously reported immunofluorescent procedure. The difficulty of two-color analysis in solid tumors using previously reported method came from the non-specific staining of resulted cells which have Fc receptors on each cell surface and hence the gaps between S- and G1- or G2M-phase were unclear. By combining proteolytic enzyme digestion, the normal cell population can be excluded easily and the gaps between these phases be clearly distinguished, therefore the results obtained by this new method seemed to be more precise. This new method can be applied for solid tumor studies and the development for clinical application is expected.
尽管已经开发出一种染色原理和同时进行流式细胞术溴脱氧尿苷(BrdU)/DNA分析的方法,但该技术在临床上很少使用,因为它不能用于实体瘤的研究。在本文中,我们描述了一种通过将蛋白水解酶消化与先前报道的免疫荧光程序相结合来克服这一问题的新方法。使用先前报道的方法对实体瘤进行双色分析的困难在于,所得细胞在每个细胞表面都有Fc受体,从而导致非特异性染色,因此S期与G1期或G2M期之间的间隙不清楚。通过结合蛋白水解酶消化,可以轻松排除正常细胞群,并清楚地区分这些阶段之间的间隙,因此这种新方法获得的结果似乎更精确。这种新方法可用于实体瘤研究,有望开发用于临床应用。