CIL:44601, Hypsiprymnodon moschatus, kidney epithelial cell. In Cell Image Library
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PtK2 cells were grown on poly-L-lysine-coated cover glasses, treated briefly with an extraction buffer containing CaCl2 and Triton X-100 to remove unpolymerized tubulin, followed by fixation for 20 min with 1% glutaraldehyde in extraction buffer. After quenching in sodium borohydride, cells were immunostained for microtubules (primary antibody mouse DM1-alpha, secondary antibody donkey anti-mouse [-Xrat] Dylight 594, red), kinetochores (primary antibody rabbit anti-CenpE, secondary antibody donkey anti-rabbit-Alexa 488, green), and chromosomes counterstained blue with Hoechst dye. Images were captured and processed using a Deltavision OMX 3D-SIM Super-Resolution microscopy system in which structured illumination provides resolution that exceeds the diffraction limit. Through-focus Z-stacks of images were obtained with a 0.125 micrometer step size and a 100x 1.4 NA UNIPLANAPO objective lens, and the z-slices subsequently processed and projected down the z-axis.
提供机构：
UC San Diego Library Digital Collections
创建时间：
2021-04-15
