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首页> 外文会议>Optical diagnostics and sensing XIV: toward point-of-care diagnostics >Dual-wavelength excitation to reduce background fluorescence for fluorescence spectroscopic quantitation of erythrocyte zinc protoporphyrin-Ⅸ and protoporphyrin-Ⅸ from whole blood and oral mucosa
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Dual-wavelength excitation to reduce background fluorescence for fluorescence spectroscopic quantitation of erythrocyte zinc protoporphyrin-Ⅸ and protoporphyrin-Ⅸ from whole blood and oral mucosa

机译：双波长激发以减少背景荧光，用于荧光光谱定量分析全血和口腔粘膜中的红细胞锌原卟啉Ⅸ和原卟啉Ⅸ

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摘要

Erythrocyte zinc protoporphyrin-Ⅸ (ZnPP) and protoporphyrin-Ⅸ (PPⅨ) accumulate in a variety of disorders that restrict or disrupt the biosynthesis of heme, including iron deficiency and various porphyrias. We describe a reagent-free spectroscopic method based on dual-wavelength excitation that can measure simultaneously both ZnPP and PPⅨ fluorescence from unwashed whole blood while virtually eliminating background fluorescence. We further aim to quantify ZnPP and PPⅨ non-invasively from the intact oral mucosa using dual-wavelength excitation to reduce the strong tissue background fluorescence while retaining the faint porphyrin fluorescence signal originating from erythrocytes. Fluorescence spectroscopic measurements were made on 35 diluted EDTA blood samples using a custom front-face fluorometer. The difference spectrum between fluorescence at 425 nm and 407 nm excitation effectively eliminated background autofluorescence while retaining the characteristic porphyrin peaks. These peaks were evaluated quantitatively and the results compared to a reference HPLC-kit method. A modified instrument using a single 1000 μm fiber for light delivery and detection was used to record fluorescence spectra from oral mucosa. For blood measurements, the ZnPP and PPⅨ fluorescence intensities from the difference spectra correlated well with the reference method (ZnPP: Spearman's rho r_s= 0.943, p＜ 0.0001; PPⅨ: r_s= 0.959, p＜ 0.0001). In difference spectra from oral mucosa, background fluorescence was reduced significantly, while porphyrin signals remained observable. The dual-wavelength excitation method evaluates quantitatively the ZnPP/heme and PPⅨ/heme ratios from unwashed whole blood, simplifying clinical laboratory measurements. The difference technique reduces the background fluorescence from measurements on oral mucosa, allowing for future non-invasive quantitation of erythrocyte ZnPP and PPⅨ.
机译：红细胞锌原卟啉-Ⅸ（ZnPP）和原卟啉-Ⅸ（PPⅨ）积聚在多种限制或破坏血红素生物合成的疾病中，包括铁缺乏症和各种卟啉症。我们描述了一种基于双波长激发的无试剂光谱方法，该方法可以同时测量未洗全血中的ZnPP和PPⅨ荧光，同时实际上消除了背景荧光。我们进一步的目标是使用双波长激发从完整的口腔粘膜中无创地定量ZnPP和PPⅨ，以减少强烈的组织背景荧光，同时保留源自红细胞的微弱卟啉荧光信号。使用定制的正面荧光计对35个稀释的EDT​​A血液样本进行荧光光谱测量。 425 nm和407 nm激发的荧光之间的差异光谱有效消除了背景自发荧光，同时保留了特征性卟啉峰。对这些峰进行定量评估，并将结果与​​参考HPLC-kit方法进行比较。使用单根1000μm光纤进行光传输和检测的改良仪器用于记录口腔粘膜的荧光光谱。对于血液测量，来自差异光谱的ZnPP和PPⅨ荧光强度与参考方法相关性很好（ZnPP：Spearman的r_s = 0.943，p ＜0.0001；PPⅨ：r_s = 0.959，p ＜0.0001）。在与口腔粘膜不同的光谱中，背景荧光显着降低，而卟啉信号仍可观察到。双波长激发法可定量评估未清洗的全血中ZnPP /血红素和PPⅨ/血红素的比率，从而简化临床实验室测量。差异技术可减少口腔黏膜测量的背景荧光，从而可用于将来对红细胞ZnPP和PPⅨ的无创定量分析。

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  • 来源
  • 会议地点 San Francisco CA(US)
  • 作者

    Georg Hennig; Michael Vogeser; Lesca M. Holdt; Christian Homann; Michael Grossmann; Herbert Stepp; Christian Gruber; Ilknur Erdogan; Stefan Hasmueller; Uwe Hasbargen; Gary M. Brittenham;

  • 作者单位

    Laser-Forschungslabor, LIFE-Zentrum, Klinikum der Universitaet Muenchen, Marchioninistr. 23, 81377 Muenchen, Germany;

    Institut fuer Laboratoriumsmedizin, Klinikum der Universitaet Muenchen, Marchioninistr. 15, 81377 Muenchen, Germany;

    Institut fuer Laboratoriumsmedizin, Klinikum der Universitaet Muenchen, Marchioninistr. 15, 81377 Muenchen, Germany;

    Laser-Forschungslabor, LIFE-Zentrum, Klinikum der Universitaet Muenchen, Marchioninistr. 23, 81377 Muenchen, Germany;

    Laser-Forschungslabor, LIFE-Zentrum, Klinikum der Universitaet Muenchen, Marchioninistr. 23, 81377 Muenchen, Germany;

    Laser-Forschungslabor, LIFE-Zentrum, Klinikum der Universitaet Muenchen, Marchioninistr. 23, 81377 Muenchen, Germany;

    Laser-Forschungslabor, LIFE-Zentrum, Klinikum der Universitaet Muenchen, Marchioninistr. 23, 81377 Muenchen, Germany;

    Klinik und Poliklinik fuer Frauenheilkunde und Geburtshilfe - Grosshadern, Klinikum der Universitaet Muenchen, Marchioninistr. 15, 81377 Muenchen, Germany;

    Klinik und Poliklinik fuer Frauenheilkunde und Geburtshilfe - Grosshadern, Klinikum der Universitaet Muenchen, Marchioninistr. 15, 81377 Muenchen, Germany;

    Klinik und Poliklinik fuer Frauenheilkunde und Geburtshilfe - Grosshadern, Klinikum der Universitaet Muenchen, Marchioninistr. 15, 81377 Muenchen, Germany;

    Division of Pediatric Hematology, Oncology and Stem Cell Transplantation, Department of Pediatrics, Columbia University College of Physicians and Surgeons, 3959 Broadway, New York, NY 10032, USA;

  • 会议组织
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    Iron deficiency; Zinc protoporphyrin Ⅸ; Protoporphyrin Ⅸ; blood; oral mucosa; non-invasive; fluorescence;

    机译：缺铁;锌原卟啉Ⅸ;原卟啉Ⅸ;血液;口腔粘膜非侵入性萤光;

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