Stout, Jane; Walczak, Claire (2021). CIL:44601, Hypsiprymnodon moschatus, kidney epithelial cell. In Cell Image Library. UC San Diego Library Digital Collections. Dataset. https://doi.org/10.6075/J0805213

Shown is a mitotic PtK2 cell at metaphase immunostained for microtubules (red) and kinetochores (green) with DNA stained blue. The image was obtained using structured illumination microscopy (SIM, Deltavision OMX system) which provides 'super-resolution' beyond the diffraction limit set by the wavelength of the illuminating light. The micrograph was the 2012 winner in the high- and super-resolution microscopy category of the GE Healthcare Life Sciences Imaging Contest, and featured in the NIGMS Biomedical Beat, the monthly digest of notable NIGMS-sponsored research.

• 2021

• Stout, Jane
• Walczak, Claire

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.

Preparation: glutaraldehyde fixed tissue
Relation to intact cell: whole mounted tissue
Item type: recorded image
Imaging mode: fluorescence microscopy; structured illumination microscopy (SIM)
Parameter imaged: fluorescence emission
Source of contrast: distribution of epitope
Visualization methods: Hoechst 33342; Alexa Fluor 488; DyLight 594
Data qualification: Processed;spatialmeasurements

• Hypsiprymnodon moschatus

• Kidney epithelial cell
• Kinetochore
• Mitotic spindle
• Nuclear chromosome

• Mitosis
• Mitotic metaphase
• PtK2

• data
• image

• No linguistic content; Not applicable

Samplenumber: 44601

Doi: https://doi.org/10.6075/J0805213

Source data

• Source Record in the Cell Image Library: https://doi.org/10.7295/W9CIL44601

Other resource

• GE Healthcare Imaging Competition announcement: https://www.ge.com/news/press-releases/winners-ge-healthcare-2012-cell-imaging-competition-announced
• GE Healthcare Imaging Competition: https://promo.gelifesciences.com/na/K12301/#.UYJa38rueRM
• NIGMS Biomedical Beat: https://biobeat.nigms.nih.gov

CC0 1.0 Universal Public Domain Dedication

Creative Commons Public Domain Dedication

Use: The person(s) who associated a work with this deed has dedicated the work to the public domain by waiving all of their rights to the work worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law.

Constraint(s) on Use: This work may be used without prior permission.

Research Data Curation Program, UC San Diego, La Jolla, 92093-0175 (https://lib.ucsd.edu/rdcp)

2022-08-12