Department of Dermatology, University Hospital Regensburg, 93042 Regensburg, Germany.
Proc Natl Acad Sci U S A. 2011 Feb 8;108(6):2432-7. doi: 10.1073/pnas.1006945108. Epub 2011 Jan 24.
Luminescence imaging of biological parameters is an emerging field in biomedical sciences. Tools to study 2D pH distribution are needed to gain new insights into complex disease processes, such as wound healing and tumor metabolism. In recent years, luminescence-based methods for pH measurement have been developed. However, for in vivo applications, especially for studies on humans, biocompatibility and reliability under varying conditions have to be ensured. Here, we present a referenced luminescent sensor for 2D high-resolution imaging of pH in vivo. The ratiometric sensing scheme is based on time-domain luminescence imaging of FITC and ruthenium(II)tris-(4,7-diphenyl-1,10-phenanthroline). To create a biocompatible 2D sensor, these dyes were bound to or incorporated into microparticles (aminocellulose and polyacrylonitrile), and particles were immobilized in polyurethane hydrogel on transparent foils. We show sensor precision and validity by conducting in vitro and in vivo experiments, and we show the versatility in imaging pH during physiological and chronic cutaneous wound healing in humans. Implementation of this technique may open vistas in wound healing, tumor biology, and other biomedical fields.
生物参数的荧光成像是生物医学科学中的一个新兴领域。为了深入了解复杂的疾病过程,如伤口愈合和肿瘤代谢,需要研究二维 pH 分布的工具。近年来,已经开发出基于荧光的 pH 测量方法。然而,对于体内应用,特别是对于人类的研究,必须确保生物相容性和在不同条件下的可靠性。在这里,我们提出了一种参考的荧光传感器,用于体内 2D 高分辨率 pH 成像。比率感应方案基于 FITC 和钌(II)三-(4,7-二苯基-1,10-菲咯啉)的时域荧光成像。为了创建一种生物相容性的 2D 传感器,将这些染料结合或掺入到微粒(氨基纤维素和聚丙烯腈)中,并将颗粒固定在透明箔上的聚氨酯水凝胶中。我们通过进行体外和体内实验来展示传感器的精度和有效性,并展示在人类生理和慢性皮肤伤口愈合过程中成像 pH 的多功能性。这项技术的实施可能会为伤口愈合、肿瘤生物学和其他生物医学领域开辟新的前景。