Ryan Lucas S, Gerberich Jeni, Haris Uroob, Nguyen Daphne, Mason Ralph P, Lippert Alexander R
Department of Chemistry, Southern Methodist University, Dallas, Texas 75275-0314, United States.
Prognostic Imaging Research Laboratory (PIRL), Pre-clinical Imaging Section, Department of Radiology, UT Southwestern Medical Center, Dallas, Texas 75390-9058, United States.
ACS Sens. 2020 Sep 25;5(9):2925-2932. doi: 10.1021/acssensors.0c01393. Epub 2020 Sep 3.
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 , 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 , which is shown and calibrated in sacrificed mouse models. Intraperitoneal injections of into live mice show high photon outputs and consistent increases in the flux ratio when measured at pH 6, 7, and 8.
生理pH值的调节对于全身和细胞的正常功能至关重要，pH值稳态的破坏可能既是疾病的原因，也是疾病的结果。鉴于此，已经开发了许多方法来监测细胞和动物中的pH值。在本研究中，我们报道了一种化学发光共振能量转移（CRET）探针，它由一个丙烯酰胺1,2 - 二氧杂环丁烷化学发光支架和一个附加的pH敏感碳荧光素荧光团组成。该探针能在6.8至8.4之间准确测量pH值，使其成为测量生物系统中pH值的可行工具。此外，其比率输出不受混杂变量的影响。pH值的定量可以使用常见的发光光谱法和先进的光学成像方法来完成。使用IVIS Spectrum，可以通过组织测量pH值，这在处死的小鼠模型中得到了展示和校准。向活小鼠腹腔注射在pH值为6、7和8时测量显示出高光子输出和通量比率的持续增加。
