Emma Kitchner, Michael Seung, Jose Chavez, Luca Ceresa, Joseph Kimball, Ignacy Gryczynski, Zygmunt Gryczynski
{"title":"荧光测量:g因子校正、魔角和观察波长的重要性。","authors":"Emma Kitchner, Michael Seung, Jose Chavez, Luca Ceresa, Joseph Kimball, Ignacy Gryczynski, Zygmunt Gryczynski","doi":"10.1088/2050-6120/ac92c5","DOIUrl":null,"url":null,"abstract":"<p><p>Excitation and emission (observation) conditions heavily impact fluorescence measurements. Both observed spectra and intensity decays (fluorescence lifetimes), when incorrectly measured, may lead to incorrect data interpretations. In this report, we discuss the role of observation conditions in steady-state and time-resolved (lifetime) fluorescence measurements. We demonstrate the importance of the correction for uneven transmissions of vertical and horizontal polarizations of emission light through the detection system. The necessity of using so-called total fluorescence intensity or intensity measured under magic angle (MA) conditions has been demonstrated for both steady-state and time-resolved fluorescence measurements. The dependence of lifetime measurements on observation (emission) wavelengths is also discussed. Two fluorophores, rhodamine 6G (R6G) and 4,4 Dimethylamino-cyano stilbene (DCS) in two solvents - ethanol and glycerol have been used in order to cover a broad range of dye polarities and solvent viscosities.</p>","PeriodicalId":18596,"journal":{"name":"Methods and Applications in Fluorescence","volume":" ","pages":""},"PeriodicalIF":2.4000,"publicationDate":"2022-09-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"1","resultStr":"{\"title\":\"Fluorescence measurements: importance of G-factor correction, magic angle, and observation wavelengths.\",\"authors\":\"Emma Kitchner, Michael Seung, Jose Chavez, Luca Ceresa, Joseph Kimball, Ignacy Gryczynski, Zygmunt Gryczynski\",\"doi\":\"10.1088/2050-6120/ac92c5\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Excitation and emission (observation) conditions heavily impact fluorescence measurements. Both observed spectra and intensity decays (fluorescence lifetimes), when incorrectly measured, may lead to incorrect data interpretations. In this report, we discuss the role of observation conditions in steady-state and time-resolved (lifetime) fluorescence measurements. We demonstrate the importance of the correction for uneven transmissions of vertical and horizontal polarizations of emission light through the detection system. The necessity of using so-called total fluorescence intensity or intensity measured under magic angle (MA) conditions has been demonstrated for both steady-state and time-resolved fluorescence measurements. The dependence of lifetime measurements on observation (emission) wavelengths is also discussed. Two fluorophores, rhodamine 6G (R6G) and 4,4 Dimethylamino-cyano stilbene (DCS) in two solvents - ethanol and glycerol have been used in order to cover a broad range of dye polarities and solvent viscosities.</p>\",\"PeriodicalId\":18596,\"journal\":{\"name\":\"Methods and Applications in Fluorescence\",\"volume\":\" \",\"pages\":\"\"},\"PeriodicalIF\":2.4000,\"publicationDate\":\"2022-09-26\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"1\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Methods and Applications in Fluorescence\",\"FirstCategoryId\":\"92\",\"ListUrlMain\":\"https://doi.org/10.1088/2050-6120/ac92c5\",\"RegionNum\":3,\"RegionCategory\":\"化学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q3\",\"JCRName\":\"CHEMISTRY, ANALYTICAL\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Methods and Applications in Fluorescence","FirstCategoryId":"92","ListUrlMain":"https://doi.org/10.1088/2050-6120/ac92c5","RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"CHEMISTRY, ANALYTICAL","Score":null,"Total":0}
Fluorescence measurements: importance of G-factor correction, magic angle, and observation wavelengths.
Excitation and emission (observation) conditions heavily impact fluorescence measurements. Both observed spectra and intensity decays (fluorescence lifetimes), when incorrectly measured, may lead to incorrect data interpretations. In this report, we discuss the role of observation conditions in steady-state and time-resolved (lifetime) fluorescence measurements. We demonstrate the importance of the correction for uneven transmissions of vertical and horizontal polarizations of emission light through the detection system. The necessity of using so-called total fluorescence intensity or intensity measured under magic angle (MA) conditions has been demonstrated for both steady-state and time-resolved fluorescence measurements. The dependence of lifetime measurements on observation (emission) wavelengths is also discussed. Two fluorophores, rhodamine 6G (R6G) and 4,4 Dimethylamino-cyano stilbene (DCS) in two solvents - ethanol and glycerol have been used in order to cover a broad range of dye polarities and solvent viscosities.
期刊介绍：
Methods and Applications in Fluorescence focuses on new developments in fluorescence spectroscopy, imaging, microscopy, fluorescent probes, labels and (nano)materials. It will feature both methods and advanced (bio)applications and accepts original research articles, reviews and technical notes.
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