Ratiometric Two-Photon Fluorescent Probe for Quantitative Detection of β-Galactosidase Activity in Senescent Cells

Abstract

We reported a ratiometric two-photon fluorescent probe (SG1) for β-galactosidase (β-gal) and its application to quantitative detection of β-gal activity during cellular senescence in live cells and in aged tissues. This probe is characterized by a significant two-photon excited fluorescence, a marked blue-to-yellow emission color change in response to β-gal, easy loading, insensitivity to pH and reactive oxygen species (ROS), high photostability, and low cytotoxicity. In addition, we show that SG1 labeling is an effective tool for quantitative detection of senescence-associated β-gal activity at the subcellular level in situ. This finding demonstrates that SG1 will find useful applications in biomedical research, including studies of cell aging.

Mitochondrial Respiratory Dysfunction Induces Claudin-1 Expression via Reactive Oxygen Species-mediated Heat Shock Factor 1 Activation, Leading to Hepatoma Cell Invasiveness

“…3F, b). This biphasic ROS generation by a subcytotoxic dose of H 2 O 2 corresponds well with a previous report (31). Interestingly, under these conditions, Cln-1 mRNA expression also showed a biphasic profile, suggesting that a certain level of ROS may be required for Cln-1 induction (Fig.…”

Progress and potential of non-inhibitory small molecule chaperones for the treatment of Gaucher disease and its implications for Parkinson disease

“…In another recent study, a ratiometric two-photon fluorescent substrate was developed for evaluation of β-galactosidase activity in live cells during cell senescence (86). The data showed that treatment of live cells with the pure product indicated a bleed-through signal of the product into the substrate channel in the two-photon microscopy assay (86).…”

“…The data showed that treatment of live cells with the pure product indicated a bleed-through signal of the product into the substrate channel in the two-photon microscopy assay (86). Such ratiometric probes are not the most ideal tools in assay development and evaluation of non-inhibitory chaperones because it will not give a definitive evaluation of the activity of chaperone-activated mutant enzyme.…”

Development of a Probe Based on Quantum Dots Embedded with Molecularly Imprinted Polymers to Detect Parathion

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