Ratiometric pH Imaging Using a 1,2-Dioxetane Chemiluminescence Resonance Energy Transfer Sensor in Live Animals
Ratiometric pH Imaging Using a 1,2-Dioxetane Chemiluminescence Resonance Energy Transfer Sensor in Live Animals
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DOI：
10.1021/acssensors.0c01393
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发表时间：
2020-09-25
期刊：
影响因子：
8.9
通讯作者：
Lippert, Alexander R.
中科院分区：
文献类型：
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作者：
Ryan, Lucas S.;Gerberich, Jeni;Lippert, Alexander R.
生理pH的调节对于适当的全身和细胞功能是不可或缺的，并且pH稳态的破坏可以是疾病的原因和结果。有鉴于此，已经开发了许多方法来监测细胞和动物中的pH值。在这项研究中，我们报告了化学发光共振能量转移（CRET）探针Ratio-pHCL-1，由丙烯酰胺1，2-二氧杂环丁烷荧光支架与附加的pH敏感的碳荧光素荧光团。该探头可精确测量6.8至8.4之间的pH值，使其成为测量生物系统中pH值的可行工具。此外，它的比率输出是独立的混杂变量。pH值的定量可以使用普通的发光光谱法和先进的光学成像方法来完成。使用IVIS Spectrum，可以通过具有Ratio-pHCL-1的组织测量pH，其在体外显示并在处死的小鼠模型中校准。当在pH 6、7和8下测量时，将Ratio-pHCL-1腹膜内注射到活小鼠中显示出高光子输出和通量比的一致增加。
Regulation of physiological pH is integral for proper whole body and cellular function, and disruptions in pH homeostasis can be both a cause and effect of disease. In light of this, many methods have been developed to monitor pH in cells and animals. In this study, we report a chemiluminescence resonance energy transfer (CRET) probe Ratio-pHCL-1, composed of an acrylamide 1,2-dioxetane chemiluminescent scaffold with an appended pH-sensitive carbofluorescein fluorophore. The probe provides an accurate measurement of pH between 6.8 and 8.4, making it a viable tool for measuring pH in biological systems. Further, its ratiometric output is independent of confounding variables. Quantification of pH can be accomplished using both common luminescence spectroscopy and advanced optical imaging methods. Using an IVIS Spectrum, pH can be measured through tissue with Ratio-pHCL-1, which is shown in vitro and calibrated in sacrificed mouse models. Intraperitoneal injections of Ratio-pHCL-1 into live mice show high photon outputs and consistent increases in the flux ratio when measured at pH 6, 7, and 8.