![Author ORCID: We display the ORCID iD icon alongside authors names on our website to acknowledge that the ORCiD has been authenticated when entered by the user. To view the users ORCiD record click the icon. [opens in a new tab]](https://chemrxiv.org/engage/assets/public/chemrxiv/images/logos/orcid.png)
Abstract
Glutathione (GSH) plays a vital role in the regulation of intracellular functions which alterations in physiological glutathione levels are associated to various diseases. Molecular imaging using fluorescent probes is a sensitive method for glutathione detection. Despite significant progress, challenges persist in the development of fluorescent probes, notably concerning potential aggregation in polar environments, quantitative GSH detection, and long-term tracking of intracellular GSH concentration. Engineered nanomaterials have shown great promise for increasing the disease diagnosis accuracy. Microchips generated by advanced microfabrication techniques can be applied in designing biomedical devices due to control over size, shape, and bioactive coatings utilization. In the current work, the synthesis and characterization of two GSH probes, Bdpy1 and Bdpy2, is reported, each offering irreversible and reversible GSH reactions, respectively. These GSH probes are immobilized on silicon oxide microchips (SOµC), micro-fabricated using photolithographic techniques, to give SOµC-Bdpy1 and SOµC-Bdpy2. Both functionalized microchips exhibited sensitivity to GSH, and, notably, the reversible SOµC-Bdpy2 showed less time dependency, making it more suitable for long-term intracellular GSH sensing. In vitro experiments in HeLa cells reveal both SOµC-Bdpy1 and SOµC-Bdpy2 were internalized in living cells, showing SOµC-Bdpy2 more reliable results (due to its less time dependency) for quantifying intracellular GSH. Remarkably, the intracellular GSH measurement was monitored by SOµC-Bdpy2 for 48h, indicating the functionalized microchips capability to detect GSH amount in different time intervals. This study introduces a promising approach for long term real-time quantification of intracellular GSH, overcoming current limitations in fluorescent probes and offering valuable insights into microchip-based sensing methodologies.
Supplementary materials
Title
Supporting Information
Description
Materials; General methods; Synthesis of Bdpy2; 1H NMR, 13C NMR, MS, and FT-IR results; Contact angle images; Fluorescence microscopy images of functionalized silicon oxide surfaces; Flow cytometry results; Fluorescence microscopy images for photobleaching assessment; Fluorescence microscopy images and fluorescence intensity profiles for time-dependent and concentration-dependent experiments; Confocal laser scanning microscopy images and ratiometric fluorescence quantification of intracellular GSH concentration; Confocal laser scanning microscopy images of internalized microchips for long-term tracking and reversibility studies.
Actions
