Intracellular thiols play an important role in various cellular functions. Changes in cellular thiol status have been linked to the pathogenesis of a variety of diseases. Monitoring of cellular thiol change in live cells through fluorescence microscopy can provide valuable information on thiol-related biochemical processes.  However, limited reagents are available for determination of thiol status in live cells through fluorescence microscopy. Bis(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)sulfane (BNOS) was developed in our laboratory as a thiol specific fluorogenic agent. The application of this reagent to monitor thiol change in live cells through fluorescence microscopy will be reported. Specifically, NCI-H226 cells were employed as a cell model.  N-Ethylmaleimide (NEM), a thiol blocking agent, was used as a reagent to modulate cellular thiols. Our data demonstrated that BNOS effectively imaged and semi-quantified cellular thiols through fluorescence microscopy. The quantification results were validated by a reported HPLC thiol assay method.

Bis(7-nitrobenzo[c][1,2,5]oxadiazol-4-yl)sulfane for thiol imaging and semi-quantification in live cells through fluorescence microscopy

244th National Meeting (2012)

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