In-situ SERS readout strategy to improve the reliability of beta-galactosidase activity assay based on X-gal staining in shortening incubation times

doi:10.1016/j.talanta.2021.122689

	In-situ SERS readout strategy to improve the reliability of beta-galactosidase activity assay based on X-gal staining in shortening incubation times
	Li, Shaofei 1,2,3; Cheng, Yizhuang 1,2; Chen, Siyu 1,2; Qin, Miao 1,2; Li, Pan 1; Yang, Liangbao 1,2
刊名	TALANTA
	2021-11-01
卷号	234
关键词	Beta-galactosidase X-gal staining SERS-active needle In-situ readout High staining background
ISSN号	0039-9140
DOI	10.1016/j.talanta.2021.122689
通讯作者	Li, Pan(lipan2011@iim.ac.cn) ; Yang, Liangbao(lbyang@iim.ac.cn)
英文摘要	Beta-galactosidase (beta-gal) activity is closed related with senescence cells and aging-associated diseases, however, the traditional readout of beta-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 beta-gal positive cells. By building the blue-white screening model and fabricating SERS-active needle sensor, the sensitive detection of beta-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 beta-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.
资助项目	National Science Foundation of China[21974142] ; Nature Science Research Project of Anhui Province[1908085QB65]
WOS关键词	SENESCENCE ; MECHANISMS ; SINGLE ; TUMOR
WOS研究方向	Chemistry
语种	英语
出版者	ELSEVIER
WOS记录号	WOS:000687342300006
资助机构	National Science Foundation of China ; Nature Science Research Project of Anhui Province
内容类型	期刊论文
源URL	[http://ir.hfcas.ac.cn:8080/handle/334002/125195]
专题	中国科学院合肥物质科学研究院
通讯作者	Li, Pan; Yang, Liangbao
作者单位	1.Chinese Acad Sci, Inst Hlth & Med Technol, Hefei Inst Phys Sci, Hefei 230031, Anhui, Peoples R China 2.Univ Sci & Technol China, Dept Chem, Hefei 230026, Anhui, Peoples R China 3.Anhui Univ, Sch Life Sci, Hefei 230601, Anhui, Peoples R China
推荐引用方式 GB/T 7714	Li, Shaofei,Cheng, Yizhuang,Chen, Siyu,et al. In-situ SERS readout strategy to improve the reliability of beta-galactosidase activity assay based on X-gal staining in shortening incubation times[J]. TALANTA,2021,234.
APA	Li, Shaofei,Cheng, Yizhuang,Chen, Siyu,Qin, Miao,Li, Pan,&Yang, Liangbao.(2021).In-situ SERS readout strategy to improve the reliability of beta-galactosidase activity assay based on X-gal staining in shortening incubation times.TALANTA,234.
MLA	Li, Shaofei,et al."In-situ SERS readout strategy to improve the reliability of beta-galactosidase activity assay based on X-gal staining in shortening incubation times".TALANTA 234(2021).