Interfacing 3D magnetic twisting cytometry with confocal fluorescence microscopy to image force responses in living cells

Yuejin Zhang, Fuxiang Wei, Yeh Chuin Poh, Qiong Jia, Junjian Chen, Junwei Chen, Junyu Luo, Wenting Yao, Wenwen Zhou, Wei Huang, Fang Yang, Yao Zhang, Ning Wang

Research output: Contribution to journalArticlepeer-review

Abstract

Cells and tissues can undergo a variety of biological and structural changes in response to mechanical forces. Only a few existing techniques are available for quantification of structural changes at high resolution in response to forces applied along different directions. 3D-magnetic twisting cytometry (3D-MTC) is a technique for applying local mechanical stresses to living cells. Here we describe a protocol for interfacing 3D-MTCTC with confocal fluorescence microscopy. In 3D-MTC, ferromagnetic beads are bound to the cell surface via surface receptors, followed by their magnetization in any desired direction. A magnetic twisting field in a different direction is then applied to generate rotational shear stresses in any desired direction. This protocol describes how to combine magnetic-field-induced mechanical stimulation with confocal fluorescence microscopy and provides an optional extension for super-resolution imaging using stimulated emission depletion (STED) nanoscopy. This technology allows for rapid real-time acquisition of a living cell’s mechanical responses to forces via specific receptors and for quantifying structural and biochemical changes in the same cell using confocal fluorescence microscopy or STED. The integrated 3D-MTC-microscopy platform takes ~20 d to construct, and the experimental procedures require ~4 d when carried out by a life sciences graduate student.

Original languageEnglish (US)
Pages (from-to)1437-1450
Number of pages14
JournalNature Protocols
Volume12
Issue number7
DOIs
StatePublished - Jul 1 2017

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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