摘要
应用荧光试剂（３３２５８Ｈｏｅｃｈｓｔ）微量测定细胞内ＤＮＡ含量，应用荧光胺（Ｆｌｕｏｒｅｓｃａｍｉｎｅ）微量测定细胞内蛋白质含量。结果：荧光试剂与ＤＮＡ的Ａ－Ｔ碱基形成复合物后，其激发波长（ＥＸ）由３２０ｎｍ移至３５０ｎｍ，发射波长（ＥＭ）由４８０ｎｍ移至４６０ｎｍ，且荧光强度大为增强。荧光胺与蛋白质反应生成强荧光化合物，在ＥＸ３９０ｎｍ，ＥＭ４７５ｎｍ条件下最低检测限为０．５μｇ／μｌ。该法操作简便，微量快速，重复性好。可用于荧光微量测定ＤＮＡ与蛋白质含量，还可用于观察细胞的增殖。
A microfluorometric method is described for the determination of intracellular DNA content with 33258 Hoechst fluorochrome as well as the determination of intracellular protein content with fluorescamine The results showed that the fluorochrome reagent presented maximal excitation and emission at 320 nm and 480 nm, respectively When the fluorochrome reagent reacted with DNA samples,these values were shifted to 350 nm and 460 nm, respectively, and the fluorescence intensity of the compound was greatly enhanced upon binding to the ATrich regions of DNA The reaction of fluorescamine with the protein formed fluorescence compound At maximal excitation 390 nm and emission 475 nm was capable of accurately quantitating as little as 05 g/l protein Microfluorometric determination was a simple and rapid method, and had a good reproducibility. It could be used to determine intracellular DNA and protein content and to observe cell proliferation
出处
《同济医科大学学报》
CSCD
1999年第3期267-268,272,共3页
Acta Universitatis Medicinae Tongji
基金
国家自然科学基金
