化学
染色
原位
孵化
假阳性悖论
检出限
拉曼光谱
可靠性(半导体)
分子生物学
色谱法
纳米技术
生物物理学
生物化学
病理
生物
材料科学
计算机科学
人工智能
光学
物理
功率(物理)
医学
有机化学
量子力学
作者
Shaofei Li,Yuanhui Cheng,Siyu Chen,Qiguang Miao,Pan Li,Liangbao Yang
出处
期刊:Talanta
[Elsevier]
日期:2021-11-01
卷期号:234: 122689-122689
被引量:1
标识
DOI:10.1016/j.talanta.2021.122689
摘要
Beta-galactosidase (β-gal) activity is closed related with senescence cells and aging-associated diseases, however, the traditional readout of β-gal activity based on X-gal staining was limited to low sensitivity in short incubation times and false positives in long incubation times. Here, we expose the potential role of insoluble X-gal hydrolysates in causing false positives by diffusion pollution depending on organic medium and then propose the in-situ Surface-enhanced Raman spectroscopy (SERS) readout strategy to identify and locate β-gal positive cells. By building the blue-white screening model and fabricating SERS-active needle sensor, the sensitive detection of β-gal has been realized with the detection limit of less than 1 nmol L−1. The in-situ SERS readout strategy is proved to be necessary and feasible to improve the reliability of X-gal staining assay through shortening the time to a few hours. Moreover, its application was also preliminarily evaluated to analyse individual cells and tissues, which showed the well consistency for judgement of β-gal activity cells at different times. Consequently, by improving reliability and reducing time consumption, this SERS readout strategy may be of great significance to promote the application of X-gal staining assay in biology and biomedicine.
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