Applicability and Limitations of Fluorescence Intensity-Based Thermometry Using a Palette of Organelle Thermometers
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 s...
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| Format: | Article |
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MDPI AG
2023-07-01
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| Series: | Chemosensors |
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| Online Access: | https://www.mdpi.com/2227-9040/11/7/375 |
| _version_ | 1827733541701550080 |
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| author | Takeru Yamazaki Xiao Liu Young-Tae Chang Satoshi Arai |
| author_facet | Takeru Yamazaki Xiao Liu Young-Tae Chang Satoshi Arai |
| author_sort | Takeru Yamazaki |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-11T01:11:47Z |
| format | Article |
| id | doaj.art-86397c1ad8bc4d24b12c4f12a92a0357 |
| institution | Directory Open Access Journal |
| issn | 2227-9040 |
| language | English |
| last_indexed | 2024-03-11T01:11:47Z |
| publishDate | 2023-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Chemosensors |
| spelling | doaj.art-86397c1ad8bc4d24b12c4f12a92a03572023-11-18T18:47:25ZengMDPI AGChemosensors2227-90402023-07-0111737510.3390/chemosensors11070375Applicability and Limitations of Fluorescence Intensity-Based Thermometry Using a Palette of Organelle ThermometersTakeru Yamazaki0Xiao Liu1Young-Tae Chang2Satoshi Arai3WPI Nano Life Science Institute, Kanazawa University, Kakuma-machi, Kanazawa 920-1192, JapanCenter for Self-Assembly and Complexity, Institute for Basic Science (IBS), Pohang 37673, Republic of KoreaCenter for Self-Assembly and Complexity, Institute for Basic Science (IBS), Pohang 37673, Republic of KoreaWPI Nano Life Science Institute, Kanazawa University, Kakuma-machi, Kanazawa 920-1192, JapanFluorescence 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.https://www.mdpi.com/2227-9040/11/7/375intracellular thermometryorganelle thermometerBODIPY rotorFLIMfluorescence |
| spellingShingle | Takeru Yamazaki Xiao Liu Young-Tae Chang Satoshi Arai Applicability and Limitations of Fluorescence Intensity-Based Thermometry Using a Palette of Organelle Thermometers Chemosensors intracellular thermometry organelle thermometer BODIPY rotor FLIM fluorescence |
| title | Applicability and Limitations of Fluorescence Intensity-Based Thermometry Using a Palette of Organelle Thermometers |
| title_full | Applicability and Limitations of Fluorescence Intensity-Based Thermometry Using a Palette of Organelle Thermometers |
| title_fullStr | Applicability and Limitations of Fluorescence Intensity-Based Thermometry Using a Palette of Organelle Thermometers |
| title_full_unstemmed | Applicability and Limitations of Fluorescence Intensity-Based Thermometry Using a Palette of Organelle Thermometers |
| title_short | Applicability and Limitations of Fluorescence Intensity-Based Thermometry Using a Palette of Organelle Thermometers |
| title_sort | applicability and limitations of fluorescence intensity based thermometry using a palette of organelle thermometers |
| topic | intracellular thermometry organelle thermometer BODIPY rotor FLIM fluorescence |
| url | https://www.mdpi.com/2227-9040/11/7/375 |
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