{"title":"Exploring the Interplay of Wavelength, Quantum Yield, and Penetration Depth in In Vivo Fluorescence Imaging.","authors":"Meital Harel, Rinat Ankri","doi":"10.1007/s10895-024-03985-2","DOIUrl":null,"url":null,"abstract":"<p><p>The intricate interplay between the irradiation wavelength, the fluorophore quantum yield (QY) and penetration depth profoundly influences the efficacy of in vivo fluorescence imaging in various applications. Understanding the complex behavior of fluorescence in vivo, specifically how variations in wavelength affect the QY of commonly used dyes and the depth of imaging is crucial for optimizing fluorescence imaging techniques, as it directly impacts the accuracy and efficiency of imaging in biological tissues. In our study, we explore these dynamics through Monte Carlo simulations conducted under conditions reflective of wide-field fluorescence imaging, examining how variations in wavelength impact the dye's QY and depth of imaging, and consequently, the fluorescence behavior. A transition in the exponential decay of the emission depth exponent is observed around the 500-600 nm range, indicating varying degrees of influence of depth on the fluorescence emission. The analysis of the fluorophore's QY reveals wavelength-dependent variations, with the most significant impact observed in the 600-700 nm range. Moreover, we continued our investigation to explore multiplexing, unveiling insights into the spacing between identical spots in multiplexing images across various depths and wavelengths.</p>","PeriodicalId":15800,"journal":{"name":"Journal of Fluorescence","volume":null,"pages":null},"PeriodicalIF":2.6000,"publicationDate":"2024-11-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Fluorescence","FirstCategoryId":"92","ListUrlMain":"https://doi.org/10.1007/s10895-024-03985-2","RegionNum":4,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
引用次数: 0
Abstract
The intricate interplay between the irradiation wavelength, the fluorophore quantum yield (QY) and penetration depth profoundly influences the efficacy of in vivo fluorescence imaging in various applications. Understanding the complex behavior of fluorescence in vivo, specifically how variations in wavelength affect the QY of commonly used dyes and the depth of imaging is crucial for optimizing fluorescence imaging techniques, as it directly impacts the accuracy and efficiency of imaging in biological tissues. In our study, we explore these dynamics through Monte Carlo simulations conducted under conditions reflective of wide-field fluorescence imaging, examining how variations in wavelength impact the dye's QY and depth of imaging, and consequently, the fluorescence behavior. A transition in the exponential decay of the emission depth exponent is observed around the 500-600 nm range, indicating varying degrees of influence of depth on the fluorescence emission. The analysis of the fluorophore's QY reveals wavelength-dependent variations, with the most significant impact observed in the 600-700 nm range. Moreover, we continued our investigation to explore multiplexing, unveiling insights into the spacing between identical spots in multiplexing images across various depths and wavelengths.
期刊介绍:
Journal of Fluorescence is an international forum for the publication of peer-reviewed original articles that advance the practice of this established spectroscopic technique. Topics covered include advances in theory/and or data analysis, studies of the photophysics of aromatic molecules, solvent, and environmental effects, development of stationary or time-resolved measurements, advances in fluorescence microscopy, imaging, photobleaching/recovery measurements, and/or phosphorescence for studies of cell biology, chemical biology and the advanced uses of fluorescence in flow cytometry/analysis, immunology, high throughput screening/drug discovery, DNA sequencing/arrays, genomics and proteomics. Typical applications might include studies of macromolecular dynamics and conformation, intracellular chemistry, and gene expression. The journal also publishes papers that describe the synthesis and characterization of new fluorophores, particularly those displaying unique sensitivities and/or optical properties. In addition to original articles, the Journal also publishes reviews, rapid communications, short communications, letters to the editor, topical news articles, and technical and design notes.