Multispectral fluorescence imaging to assess pH in biological specimens.

摘要: Simple, quantitative assays to measure pH in tissue could improve the study of complicated biologi- cal processes and diseases such as cancer. We evaluated multispectral fluorescence imaging (MSFI) quantify extracellular (pHe) dye-perfused, surgically-resected tumor specimens with commercially available instru- mentation. Utilizing a water-soluble organic dye pH-dependent emission (SNARF-4F), we used standard fluorimetry quantitatively assess properties function pH. By conducting these studies within spectroscopic constraints imposed by appropriate filter set supplied system, determined that correction deprotonated was necessary for accurate determination due suboptimal excitation. Subsequently, employing fluorimetry-derived factor (CF), MSFI data sets aqueous solutions tissuelike phantoms be spectrally un- mixed accurately equilibrium concentrations protonated (HA) (A − ) thus determine solution Finally, explored feasibility high-resolution pHe mapping human colorectal cancer cell-line xenografts. Data presented suggest is suitable ex vivo dye-perfused tissue, potentially enabling measurement across variety preclinical models disease. C 2011 Society Photo-Optical Instrumentation Engineers (SPIE). (DOI: 10.1117/1.3533264)