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Updated: Aug 28, 2025

Detection of Total Reactive Oxygen Species in Adherent Cells by 2',7'-Dichlorodihydrofluorescein Diacetate Staining
Published on: June 23, 2020
Carbon dot based nucleus targeted fluorescence imaging and detection of nuclear hydrogen peroxide in living cells
Kabyashree Phukan1, Ritwick Ranjan Sarma1, Somarani Dash2
1Material Nanochemistry Laboratory, Physical Sciences Division India devasish@iasst.gov.in +91 3612270095.
Abstract:
Investigation of the intracellular generation of H2O2, one of the most important reactive oxygen species (ROS), is crucial for preventing various diseases since it is closely linked with different physiological and complex cell signaling pathways. Despite the development of various fluorescent probes, the majority of the fluorescent probes cannot move across the nuclear membrane. However, detection of the nuclear level of H2O2 is very important since it can directly cause oxidative DNA damage which ultimately leads to various diseases. Therefore, in this study, p-phenylenediamine based carbon quantum dots (B-PPD CDs) have been synthesized and integrated with 4-formylbenzeneboronic acid as a doping agent for the detection of H2O2. The detection mechanism showed that, upon exposure to H2O2, the fluorescence of the B-PPD CDs was immediately quenched. Further investigation has been done in the in vitro RAW 264.7 cell line by both exogenous and endogenous exposure of H2O2 to demonstrate the feasibility of the method. It is shown successfully that the exogenous presence and endogenous generation of H2O2 in RAW 264.7 cells can be detected using B-PPD CDs. The limit of detection (LOD) was determined to be 0.242 μM. The development of such imaging probes using carbon quantum dots will lead to live-cell imaging as well as ROS detection.

