Debaditya Mukhopadhyay, Alexei Arnaoutov, Mary Dasso (2011) CIL:14008, Homo sapiens. CIL. Dataset

Hela cells were treated with anti-SENP6 siRNA and synchronized by DTB for 48 hours. Cells were fixed and stained for kinetochore proteins CENP-E (green, secondary Ab conjugated with Alexa Fluor 488) Cells were simultaneously stained for centromeres (CREST, red, secondary Ab conjugated with Alexa Fluor 568) and Hoechst 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).

Image Reference: PMID 20212317

人肺腺癌细胞系 Hela 经抗 SEPN6 siRNA 处理，并通过 DTB 方法同步培养 48 小时。细胞经固定后，采用针对着丝粒蛋白 CENP-E（绿色，经 Alexa Fluor 488 标记的二抗）进行染色。同时，细胞亦进行着丝粒（CREST，红色，经 Alexa Fluor 568 标记的二抗）和 Hoechst 33342（蓝色）的染色。在室温下，使用共聚焦显微镜（LSM510 Meta；卡尔·蔡司公司）及 100× Plan-Apochromat 物镜进行荧光显微镜观察。检测 Alexa Fluor 568 标记的抗体时，采用 543 nm 的 HeNe 激光（输出功率 5 mW；检测波长 LP560 nm）。分析 Alexa Fluor 488 标记的抗体时，使用氩激光的 488 nm 波长（标称输出功率 25 mW；检测波长 BP 505–530 nm）。Hoechst 33258 图像的捕捉利用离子激光的 364 nm 波长（Enterprise II ML UV；Coherent 公司；标称输出功率 80 mW；检测波长 BP 385–470 nm）。图像参考文献：PMID 20212317