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Functionalized CdSe/ZnS Quantum Dots for Intracellular pH Measurements by Fluorescence Lifetime Imaging Microscopy.
ACS Sensors ( IF 8.2 ) Pub Date : 2020-06-18 , DOI: 10.1021/acssensors.0c00719
Pedro J Pacheco-Liñán 1 , Iván Bravo 1, 2 , María L Nueda 2, 3 , José Albaladejo 4 , Andrés Garzón-Ruiz 1
Affiliation  

pH is an important biomarker for many human diseases and great efforts are being made to develop new pH probes for bioimaging and biomedical applications. Here, the use of three different CdSe/ZnS QDs, functionalized with d-penicillamine and small peptides, as pH probes for fluorescence lifetime imaging microscopy (FLIM) is investigated. The fluorescence pH sensitivity of the nanoparticles is analyzed in different experimental media: aqueous solution, synthetic intracellular medium, and mesenchymal C3H10T1/2 and tumoral SK-MEL-2 cell lines. Different experiments along with theoretical calculations are conducted to unravel the mechanisms causing pH sensitivity of the nanoparticles and the effect of the length and composition of the peripheral branches on their photophysical properties. Absolute intracellular pH values measured in live cells with FLIM using a fluorescent probe based on a QD are reported here for the first time (intracellular pH values of 7.0 and 7.1 for C3H10T1/2 and SK-MEL-2 cells, respectively). These fluorescent nanoprobes can also be used to distinguish between different types of cells in cocultures on the basis of their different fluorescence lifetimes in dissimilar intracellular environments.

中文翻译:

功能化的CdSe / ZnS量子点,用于通过荧光寿命成像显微镜测量细胞内pH。

pH是许多人类疾病的重要生物标志物,并且正在努力开发用于生物成像和生物医学应用的新型pH探针。在这里,使用三种不同的CdSe / ZnS QD,并用d-青霉胺和小肽,作为用于荧光寿命成像显微镜(FLIM)的pH探针,已得到研究。在不同的实验介质(水溶液,合成的细胞内介质,间充质C3H10T1 / 2和肿瘤SK-MEL-2细胞系)中分析了纳米颗粒的荧光pH敏感性。进行了不同的实验以及理论计算,以阐明引起纳米粒子pH敏感性的机制以及外围分支的长度和组成对其光物理性质的影响。首次报道了使用基于QD的荧光探针在FLIM的活细胞中测得的细胞内绝对pH值(C3H10T1 / 2和SK-MEL-2细胞的细胞内pH值分别为7.0和7.1)。
更新日期:2020-07-24
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