Live-cell imaging of LLC-PK1 cells microtubule dynamics

DOI10.5281/zenodo.6850280Zenodo6850280MaRDI QIDQ6690328FDOQ6690328

Dataset published at Zenodo repository.

A Linares

Publication date: 17 July 2022

Copyright license: Creative Commons Attribution 4.0 International

Description

Stable LLC-PK1 cell line (ATCC:CL-101) was generated and provided by Michael W. Davidson [1,2]. Using a Zeiss Celldiscoverer 7 microscope with a 100X/1.47 NA oil-immersion objective (Plan-Apochromat, Zeiss) and a sCMOS sensor (Hamamatsu, ORCA-Fusion, C15440-20UP), cultured LLC-PK1 cells expressing mEmerald-EB3 were imaged with a 43 nm pixel size. Exposure time was 100 ms, with each frame being captured every two seconds. During acquisition, temperature and CO2 control was set to 37C and 5%, respectively. mEmerald-EB3 was excited using a 488 nm laser at 1% power, with a FITC filter for fluorescence collection. Acquisition software ZEN 3.2 (blue edition) was used for the imaging protocol. 1.Rizzo, M. A., Davidson, M. W., Piston, D. W. (2009). Fluorescent protein tracking and detection: fluorescent protein structure and color variants. Cold Spring Harbor Protocols, 2009(12), pdb-top63. 2.Huang, F., Hartwich, T., Rivera-Molina, F. et al. Video-rate nanoscopy using sCMOS cameraspecific single-molecule localization algorithms. Nat Methods 10, 653658 (2013).

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