Myers, Kenneth; Waterman, Clare M. (2021). CIL:7437, Homo sapiens, endothelial cell, primary cell line cell. In Cell Image Library. UC San Diego Library Digital Collections. Dataset. https://doi.org/10.6075/J03777SN

Sub-confluent, interphase Human Umbilical Vein Endothelial Cells (HUVECs) transfected with GFP-WT-MCAK (Mitotic Centromere Associated Kinesin) and grown in culture for 8 hours. Cells were fixed in PHEM buffer containing 4% paraformaldehyde, 0.05% gluteraldehyde, and 0.2% Triton X-100. Cells were then labeled with mouse anti-tubulin detected by goat anti-mouse Alexa 568 antibody.

Cells were imaged on a spinning disk confocal microscope using a 60x 1.4 NA oil immersion objective lens on a Nikon Eclipse Ti equipped with Perfect Focus System and a Yokogawa CSU-X spinning disk confocal scan head equipped with a multi-bandpass dichromatic mirror (Semrock; Rochester, NY) and bandpass filters (Chroma; Rockingham, VT) in an electronic filterwheel for selection of GFP or Texas red emission. Excitation light provided by a custom-built laser combiner module (modification of LMM-3, Spectral Applied Research; Richmond Hill, Ontario, Canada) containing 500mW solid state lasers (488 nm: Coherent; 561 nm: MPB Communications; Montreal, Quebec) that were shuttered with electronic shutters and attenuated and/or directed to a fiber-coupled output port with an AOTF (Neos Technologies, Melbourne, FL) and directed to the confocal scan-head via a single-mode optical fiber (Oz Optics, Ottawa, Ontario, Canada). A Coolsnap HQ2 camera captured images with 200ms exposure @ 488nm and 400ms exposure @ 560nm.

• 2021

• Myers, Kenneth
• Waterman, Clare M.

Preparation: formaldehyde fixed tissue; glutaraldehyde fixed tissue; detergent permeabilized
Relation to intact cell: dispersed cells in vitro
Item type: recorded image
Imaging mode: spinning disk confocal microscopy
Parameter imaged: fluorescence emission
Source of contrast: differences in adsorption or binding of stain
Visualization methods: EGFP; primary antibody plus labeled secondary antibody; Alexa Fluor 568
Processing history: unprocessed raw data

• Homo sapiens

• Endothelial cell
• Kinesin complex
• Microtubule
• Primary cell line cell

• Protein localization to chromosome, centromeric region

• data
• image

• No linguistic content; Not applicable

Samplenumber: 7437

Doi: https://doi.org/10.6075/J03777SN

Source data

• Source Record in the Cell Image Library: https://doi.org/10.7295/W9CIL7437

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

2022-08-12