Temperature imaging using a cationic linear fluorescent polymeric thermometer and fluorescence lifetime imaging microscopy View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2019-03-22

AUTHORS

Noriko Inada, Nanaho Fukuda, Teruyuki Hayashi, Seiichi Uchiyama

ABSTRACT

Temperature is one of the most important of the physiological parameters that determine the biological status of living organisms. However, intracellular temperature was not imaged at the single-cell level until recently because of the lack of a molecular thermometer that can be applied to living cells. We have recently developed a method for imaging intracellular temperature using a cationic linear fluorescent polymeric thermometer (FPT) and fluorescence lifetime imaging microscopy (FLIM). The cationic linear FPT exhibits cell permeability in various mammalian cell lines and yeast cells, entering live cells within 10 min of incubation. Intracellular thermometry using the cationic linear FPT and FLIM can be used to image temperature with high temperature resolution (0.3-1.29 °C within a temperature range of 25-35 °C). The diffuse intracellular localization of the cationic linear FPT allows a high spatial resolution (i.e., the light microscope's diffraction limit, 200 nm), enabling the detection of temperature distributions at the subcellular level. This protocol, including the construction of a calibration curve and intracellular temperature imaging, requires ~14 h. Experience in handling cultured mammalian cells and use of a confocal laser-scanning microscope (CLSM) is required. More... »

PAGES

1293-1321

References to SciGraph publications

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/s41596-019-0145-7

DOI

http://dx.doi.org/10.1038/s41596-019-0145-7

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1112949975

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/30903107


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