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Applicability and Limitations of Fluorescence Intensity-Based Thermometry Using a Palette of Organelle Thermometers

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Title
Applicability and Limitations of Fluorescence Intensity-Based Thermometry Using a Palette of Organelle Thermometers
Author(s)
Yamazaki, Takeru; Xiao Liu; Young-Tae Chang; Arai, Satoshi
Publication Date
2023-07
Journal
CHEMOSENSORS, v.11, no.7
Publisher
MDPI
Abstract
Fluorescence thermometry is a microscopy technique in which a fluorescent temperature sensor records temperature changes as alterations of fluorescence signals. Fluorescence lifetime imaging (FLIM) is a promising method for quantitative analysis of intracellular temperature. Recently, we developed small-molecule thermometers, termed Organelle Thermo Greens, that target various organelles and achieved quantitative temperature mapping using FLIM. Despite its highly quantitative nature, FLIM-based thermometry cannot be used widely due to expensive instrumentation. Here, we investigated the applicability and limitations of fluorescence intensity (FI)-based analysis, which is more commonly used than FLIM-based thermometry. Temperature gradients generated by artificial heat sources and physiological heat produced by brown adipocytes were visualized using FI- and FLIM-based thermometry. By comparing the two thermometry techniques, we examined how the shapes of organelles and cells affect the accuracy of the temperature measurements. Based on the results, we concluded that FI-based thermometry could be used for "qualitative", rather than quantitative, thermometry under the limited condition that the shape change and the dye leakage from the target organelle were not critical.
URI
https://pr.ibs.re.kr/handle/8788114/13717
DOI
10.3390/chemosensors11070375
ISSN
2227-9040
Appears in Collections:
Center for Self-assembly and Complexity(복잡계 자기조립 연구단) > 1. Journal Papers (저널논문)
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