National Institute of Laser Enhanced Sciences, Ca iro University, Giza 12613, Egypt; Egypt Nanotechnology Center, Cairo University, El-Sheikh Zayed 12588, Egypt.
National Institute of Laser Enhanced Sciences, Ca iro University, Giza 12613, Egypt.
Biosens Bioelectron. 2017 Dec 15;98:415-420. doi: 10.1016/j.bios.2017.07.020. Epub 2017 Jul 8.
Cytochrome c (Cyt c) is commonly used as intrinsic biomarker for several characteristics of the cell such as respiration, energy level and apoptosis. In the present study a simple colorimetric sensor should be developed and tested for the real-time detection of Cyt c in living cells. We synthesized cadmium telluride quantum dots (CdTe QDs) capped with thioglycolic acid (TGA) as a fluorometric Cyt c nanosensor. The synthesized TGA/CdTe QDs nanosensor was characterized by Fourier transform infrared spectroscopy, transmission electron microscopy, and absorption as well as fluorescence spectrophotometry. We investigated the developed TGA/CdTe QDs sensor with regard to its applicability in the fluorometric detection of Cyt c. Results showed that the TGA/CdTe QDs could be used as a sensitive fluorescence probe for the quantification of different concentrations of Cyt c ranging from 0.5 - 2.5μM. Increased binding of QDs to Cyt c results in decreasing fluorescence. The fluorescence of the QDs is inversely correlated to the Cyt c concentration. Based on these data, a standard curve up to 2.5μM Cyt c was established. Moreover, the developed nanosensor was applied in different concentrations on primary human dermal fibroblasts. Results showed that TGA/CdTe QDs were taken up by cells and could be visualized by fluorescence microscopy. Quantification of Cyt c within living cells via QDs is, however, influenced by various factors such as cell damage, QD aggregation or the level of reactive oxygen species, which have to be taken into account.
细胞色素 c(Cyt c)通常被用作细胞呼吸、能量水平和细胞凋亡等特性的固有生物标志物。在本研究中,应开发一种简单的比色传感器,并对其进行测试,以实时检测活细胞中的 Cyt c。我们合成了巯基乙酸(TGA)包覆的碲化镉量子点(CdTe QDs)作为荧光 Cyt c 纳米传感器。通过傅里叶变换红外光谱、透射电子显微镜、吸收和荧光分光光度法对合成的 TGA/CdTe QDs 纳米传感器进行了表征。我们研究了开发的 TGA/CdTe QDs 传感器在荧光检测 Cyt c 方面的适用性。结果表明,TGA/CdTe QDs 可作为一种灵敏的荧光探针,用于定量不同浓度的 Cyt c,范围为 0.5-2.5μM。QDs 与 Cyt c 的结合增加导致荧光降低。QDs 的荧光与 Cyt c 的浓度呈反比。基于这些数据,建立了高达 2.5μM Cyt c 的标准曲线。此外,该纳米传感器已应用于不同浓度的原代人真皮成纤维细胞。结果表明,TGA/CdTe QDs 被细胞摄取,并可通过荧光显微镜观察到。然而,通过 QDs 对活细胞内 Cyt c 的定量受到各种因素的影响,例如细胞损伤、QD 聚集或活性氧水平,这些因素都需要考虑在内。