GFP fluorescence peak fraction analysis based nanothermometer for the assessment of exothermal mitochondria activity in live cells

Nanothermometry methods with intracellular sensitivities have the potential to make important contributions to fundamental cell biology and medical fields, as temperature is a relevant physical parameter for molecular reactions to occur inside the cells and changes of local temperature are well iden...

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Bibliographic Details
Published in:Scientific reports 2019-05, Vol.9 (1), p.7535-11, Article 7535
Main Authors: Savchuk, Oleksandr A., Silvestre, Oscar F., Adão, Ricardo M. R., Nieder, Jana B.
Format: Article
Language:English
Subjects:
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Biosensing Techniques - instrumentation
Biosensing Techniques - methods
Cell Line, Tumor
Green Fluorescent Proteins - chemistry
HeLa Cells
Humanities and Social Sciences
Humans
Intracellular
Mitochondria
Mitochondria - metabolism
multidisciplinary
Science
Science (multidisciplinary)
Spatial discrimination
Temperature
Temperature effects
Thermometers
Thermometry - instrumentation
Thermometry - methods
Thermosensing
Transfection
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Summary:Nanothermometry methods with intracellular sensitivities have the potential to make important contributions to fundamental cell biology and medical fields, as temperature is a relevant physical parameter for molecular reactions to occur inside the cells and changes of local temperature are well identified therapeutic strategies. Here we show how the GFP can be used to assess temperature-based on a novel fluorescence peak fraction method. Further, we use standard GFP transfection reagents to assess temperature intracellularly in HeLa cells expressing GFP in the mitochondria. High thermal resolution and sensitivity of around 0.26% °C −1 and 2.5% °C −1 , were achieved for wt-GFP in solution and emGFP-Mito within the cell, respectively. We demonstrate that the GFP-based nanothermometer is suited to directly follow the temperature changes induced by a chemical uncoupler reagent that acts on the mitochondria. The spatial resolution allows distinguishing local heating variations within the different cellular compartments. Our discovery may lead to establishing intracellular nanothermometry as a standard method applicable to the wide range of live cells able to express GFP.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-019-44023-7