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Method Article
Martin Mistrik
Laboratory of Genome Integrity, Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacky University, Olomouc, Czech Republic
Corresponding Author
Jiri Bartek
Danish Cancer Society Research Center, Copenhagen, Denmark
Corresponding Author
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Despite proteotoxic stress and heat shock are implicated in diverse pathologies, currently no methodology to inflict defined, subcellular thermal damage exists. Here, we present a protocol for such a single-cell method compatible with laser-scanning microscopes, adopting the plasmon resonance principle. The method is based on modified microscopic cell culture plates, pre-coated by a layer of anisotropic silver NPs allowing excitation through targeted irradiation by conventional lasers used in the laser scanning microscopes and allowing controllable heating. Dose-defined heat causes protein damage in subcellular compartments, rapid heat-shock chaperones recruitment and stress signalling, thereby allowing unprecedented spatiotemporal analysis of thermal damage with broad applicability in biomedicine.
Keywords
plasmon, silver nanoparticles, heat shock response, thermal damage, protein aggregation, confocal microscopy, laser micro-irradiation
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An open repository of community-contributed protocols sponsored by Nature Research.
Learn more about Protocol Exchange
Method Article
Martin Mistrik
Laboratory of Genome Integrity, Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacky University, Olomouc, Czech Republic
Corresponding Author
Jiri Bartek
Danish Cancer Society Research Center, Copenhagen, Denmark
Corresponding Author
Version History
CURRENT STATUS: Posted
Version 1
Posted 01 Apr, 2021
No community comments so far
Metrics
Comments: 0
HTML Views: ...
Subject Areas
Biological techniques
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Despite proteotoxic stress and heat shock are implicated in diverse pathologies, currently no methodology to inflict defined, subcellular thermal damage exists. Here, we present a protocol for such a single-cell method compatible with laser-scanning microscopes, adopting the plasmon resonance principle. The method is based on modified microscopic cell culture plates, pre-coated by a layer of anisotropic silver NPs allowing excitation through targeted irradiation by conventional lasers used in the laser scanning microscopes and allowing controllable heating. Dose-defined heat causes protein damage in subcellular compartments, rapid heat-shock chaperones recruitment and stress signalling, thereby allowing unprecedented spatiotemporal analysis of thermal damage with broad applicability in biomedicine.
Figure 1
Figure 2
Figure 3
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