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二碘苯酚吲哚和4',6-二脒基-2-苯基吲哚:荧光显带,褪色可忽略不计。

DIPI and DAPI: fluorescence banding with only negliglible fading.

作者信息

Schnedl W, Mikelsaar A V, Breitenbach M, Dann O

出版信息

Hum Genet. 1977 Apr 15;36(2):167-72. doi: 10.1007/BF00273255.

Abstract

DIPI and DAPI produce distinct fluorescent bands in human chromosomes similar to quinacrine banding patterns. Additionally, the AT rich secondary constrictions in the chromosomes Nos. 1, 9 and 16 are brightly fluorescent. On the other hand the brilliantly fluorescent regions after staining with quinacrine mustard in the chromosomes Nos. 3 and 4, satellites and some other regions in the acrocentric chromosomes are less striking. The distal part of the Y, however, is clearly discernible. Thus DIPI and DAPI seem to be strictly AT specific fluorochromes like Hoechst 33258. In interphase nuclei the Y chromosome can be identified. However, quinacrines are superior for Y-body analysis in buccal, hair cell and sperm smears. BrdU labeled chromatids show reduced fluorescence intensity. The difference, however, is less apparent than after staining with Hoechst 33 258. DAPI and especially DIPI are highly resistant to UV-irradiation; there is almost no fading within 30 min when using DIPI. Moreover, fluorescence intensity is stronger than in quinicrines. When photographing, exposure times may be reduced to about one quarter compared to quinacrine mustard.

摘要

二碘化丙锭(DIPI)和4,6-二脒基-2-苯基吲哚(DAPI)在人类染色体中产生与喹吖因显带模式相似的独特荧光带。此外,1号、9号和16号染色体中富含AT的次缢痕发出明亮荧光。另一方面,用喹吖因氮芥染色后,3号和4号染色体中荧光强烈的区域、近端着丝粒染色体中的随体及其他一些区域的荧光不太明显。然而,Y染色体的远端部分清晰可辨。因此,DIPI和DAPI似乎像Hoechst 33258一样是严格的AT特异性荧光染料。在间期核中可以识别出Y染色体。然而,在颊黏膜、毛细胞和精子涂片的Y小体分析中,喹吖因更具优势。用5-溴脱氧尿嘧啶核苷(BrdU)标记的染色单体显示出降低的荧光强度。然而,这种差异不如用Hoechst 33258染色后明显。DAPI尤其是DIPI对紫外线照射具有高度抗性;使用DIPI时,30分钟内几乎没有褪色。此外,荧光强度比喹吖因更强。拍照时,与喹吖因氮芥相比,曝光时间可减少至约四分之一。

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