Abstract：
: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.
journal_name
Talantajournal_title
Talantaauthors
Li S,Cheng Y,Chen S,Qin M,Li P,Yang Ldoi
10.1016/j.talanta.2021.122689keywords:
["Beta-galactosidase","High staining background","In-situ readout","SERS-active needle","X-gal staining"]subject
Has Abstractpub_date
2021-11-01 00:00:00pages
122689eissn
0039-9140issn
1873-3573pii
S0039-9140(21)00610-Xjournal_volume
234pub_type
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