Badugu Ramachandram, Jeng Bennie H, Reece E Albert, Lakowicz Joseph R
Center for Fluorescence Spectroscopy, Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, 725 W. Lombard Street, Baltimore, MD 21201, USA.
Department of Ophthalmology and Visual Sciences, University of Maryland School of Medicine, 419 W. Redwood Street, Suite 420, Baltimore, MD 21201, USA.
Anal Biochem. 2018 Feb 1;542:84-94. doi: 10.1016/j.ab.2017.11.014. Epub 2017 Nov 26.
Dry eye disease (DED) affects millions of individuals in the United States and worldwide, and the incidence is increasing with an aging population. There is widespread agreement that the measurement of total tear osmolarity is the most reliable test, but this procedure provides only the total ionic strength and does not provide the concentration of each ionic species in tears. Here, we describe an approach to determine the individual ion concentrations in tears using modern silicone hydrogel (SiHG) contact lenses. We made pH (or HO, hydronium cation,/OH, hydroxyl ion) and chloride ion (two of the important electrolytes in tear fluid) sensitive SiHG contact lenses. We attached hydrophobic C18 chains to water-soluble fluorescent probes for pH and chloride. The resulting hydrophobic ion sensitive fluorophores (H-ISF) bind strongly to SiHG lenses and could not be washed out with aqueous solutions. Both H-ISFs provide measurements which are independent of total intensity by use of wavelength-ratiometric measurements for pH or lifetime-based sensing for chloride. Our approach can be extended to fabricate a contact lens which provides measurements of the six dominant ionic species in tears. This capability will be valuable for research into the biochemical processes causing DED, which may improve the ability to diagnose the various types of DED.
在美国及全球范围内,数百万的人都患有干眼病(DED),并且随着人口老龄化,其发病率正在上升。人们普遍认为,测量总泪液渗透压是最可靠的检测方法,但该方法仅能提供总离子强度,无法给出泪液中每种离子成分的浓度。在此,我们描述了一种使用现代硅水凝胶(SiHG)隐形眼镜来测定泪液中各离子浓度的方法。我们制作了对pH值(或水合氢离子/H⁺、氢氧根离子/OH⁻)和氯离子(泪液中的两种重要电解质)敏感的SiHG隐形眼镜。我们将疏水的C18链连接到用于检测pH值和氯离子的水溶性荧光探针上。由此产生的疏水离子敏感荧光团(H-ISF)与SiHG镜片紧密结合,且不会被水溶液冲洗掉。通过对pH值进行波长比率测量或对氯离子进行基于寿命的传感,这两种H-ISF都能提供与总强度无关的测量结果。我们的方法可以扩展到制造一种能够测量泪液中六种主要离子成分的隐形眼镜。这种能力对于研究导致干眼病的生化过程将具有重要价值,这可能会提高诊断各种类型干眼病的能力。