Debaditya Mukhopadhyay, Alexei Arnaoutov, Mary Dasso (2011) CIL:13994, Homo sapiens. CIL. Dataset

This image is part of Fig 2B from the reference article PMID 20212317. HeLa cells were treated with anti-SENP6 siRNA and double thymidine block for 48 h. Cells were fixed and stained for centromeres (CREST, red, Alexa Fluor 568 conjugated secondary Ab), Bub1 (green, Alexa Fluor 488 conjugated secondary Ab) and Hoechest 33342 (blue).

Fluorescence microscopy was performed at RT on a confocal microscope (LSM510 Meta; Carl Zeiss, Inc.) equipped with a 100× Plan-Apochromat objective. A 543 nm HeNe laser (5 mW output; detection LP560 nm) was used for detection of Alexa Fluor 568–labeled antibodies. The 488nm line of an Argon laser (25 mW nominal output; detection BP 505–530 nm) was used for analysis of Alexa Fluor 488–labeled antibodies. Hoechst 33258 images were captured using the 364nm line of an ion laser (Enterprise II ML UV; Coherent, Inc.; 80 mW nominal output; detection BP 385–470 nm).

该图像摘自参考文献文章图2B（PMID 20212317）。HeLa细胞经过抗SENP6 siRNA和双胸苷酸阻断处理48小时。细胞经固定后，以CREST（红色，Alexa Fluor 568偶联的二级抗体）染色进行着丝粒标记，以Alexa Fluor 488偶联的二级抗体对Bub1（绿色）进行标记，并以Hoechest 33342（蓝色）进行染色。在室温下，使用配置有100× Plan-Apochromat物镜的共聚焦显微镜（LSM510 Meta；卡尔·蔡司公司）进行荧光显微镜观察。采用543 nm的HeNe激光（输出功率为5 mW；检测波长为LP560 nm）用于检测Alexa Fluor 568标记的抗体。使用氩激光的488 nm波长（标称输出功率为25 mW；检测波长为BP 505–530 nm）分析Alexa Fluor 488标记的抗体。使用离子激光的364 nm波长（Coherent公司Enterprise II ML UV型号；标称输出功率为80 mW；检测波长为BP 385–470 nm）捕获Hoechst 33258图像。