A novel near-infrared fluorescent probe for sensitive detection of β-galactosidase in living cells
Open accessAbstract
A novel near-infrared fluorescent probe for β-galactosidase has been developed based on a hemicyanine skeleton, which is conjugated with a d-galactose residue via a glycosidic bond. The probe serves as a substrate of β-galactosidase and displays rapid and sensitive turn-on fluorescent responses to β-galactosidase in aqueous solution. A 12.8-fold enhancement of fluorescence intensity at 703 nm was observed after incubation of 10 nM of β-galactosidase with 5 μM probe for 10 min. The probe can sensitively detect as little as 0.1 nM of β-galactosidase and shows linear responses to the enzyme concentration below 1.4 nM. The kinetic study showed that the probe has high binding affinity to β-galactosidase with K m = 3.6 μM. The probe was used to detect β-galactosidase in living cells by employing the premature cell senescence model. The probe exhibited strong fluorescent signals in senescent cells but not in normal cells, which demonstrates that the probe is able to detect the endogenous senescence-associated β-galactosidase in living cells.
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The FACS method based on SPiDER-β-gal labels all SA-β-gal+ cells.
“we established a fluorescence-activated cell sorting (FACS) method based on the fluorescent probe SPiDER-β-gal (hereafter, SPiDER; similar to the C12FDG fluorescent probe)25,26, which labels all SA-β-gal+ cells (Extended Data Fig. 1d).”
From Senescence atlas reveals an aged-like inflamed niche that blunts muscle regeneration (2023) · cites this paper as [25]
Sections 2.2 and 3.5 describe fluorescence microscopy of senescent human fibroblasts using Gal-Pro, not SPiDER-β-gal or FACS. The copy does not address whether a SPiDER-based sorting method labels all SA-β-gal-positive cells (searched: SPiDER, FACS, C12FDG, flow cytometry). (LLM, conf=0.78)
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- Authors
- Zhang, J., Li, C., Dutta, C., Fang, M., Zhang, S., Tiwari, A., Werner, T., Luo, F.-T., & Liu, H.
- Affiliations
- Michigan Technological University (US)
- Institute of Chemistry, Academia Sinica (TW)
- Corresponding author
- Tiwari, Ashutosh
- Werner, Thomas
- Luo, Fen-Tair
- Liu, Haiying
- ORCID
- Venue
- Analytica Chimica Acta
- Publisher
- Elsevier BV
- Published
- 2017-05
- Volume
- 968
- Pages
- 97-104
- ISSN
- 0003-2670 (ISSN-L)
- PMC
- PMC5455349
- OpenAlex
- W2594193979
- Cited by
- 101
- References
- 28
- Citations by year
- 2026: 3 · 2025: 8 · 2024: 8 · 2023: 9 · 2022: 16 · 2021: 15 · 2020: 14 · 2019: 10 · 2018: 17 · 2017: 1
- FWCI
- 5.66
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- Analytica Chimica Acta (acceptedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Digital Commons - Michigan Tech (Michigan Technological University) (submittedVersion)
- Funders
- National Institute of General Medical Sciences of the National Institutes of Health (R15GM114751)
- National Science Foundation (1048655)
- Topics
- Physical Sciences › Chemistry › Spectroscopy › Molecular Sensors and Ion Detection
- Physical Sciences › Materials Science › Materials Chemistry › Luminescence and Fluorescent Materials
- Health Sciences › Medicine › Physiology › Lysosomal Storage Disorders Research
- Keywords
- beta-galactosidase, near-infrared fluorescent probe, hemicyanine, galactose, turn-on fluorescence, enzyme activity assay, live-cell imaging, senescence-associated β-galactosidase, glycosidic linkage
- MeSH terms
- beta-Galactosidase, Cells, Cultured, Fibroblasts, Fluorescent Dyes, Humans, Spectroscopy, Near-Infrared
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