A TCF-Based Carbon Monoxide NIR-Probe without the Interference of BSA and Its Application in Living Cells

Abstract

As toxic gaseous pollution, carbon monoxide (CO) plays an essential role in many pathological and physiological processes, well-known as the third gasotransmitter. Owning to the reducibility of CO, the Pd0-mediated Tsuji-Trost reaction has drawn much attention in CO detection in vitro and in vivo, using allyl ester and allyl ether caged fluorophores as probes and PdCl2 as co-probes. Because of its higher decaging reactivity than allyl ether in the Pd0-mediated Tsuji-Trost reaction, the allyl ester group is more popular in CO probe design. However, during the application of allyl ester caged probes, it was found that bovine serum albumin (BSA) in the fetal bovine serum (FBS), an irreplaceable nutrient in cell culture media, could hydrolyze the allyl ester bond, and thus give erroneous imaging results. In this work, dicyanomethylenedihydrofuran (TCF) and dicyanoisophorone (DCI) were selected as electron acceptors for constructing near-infrared-emission fluorophores with electron donor phenolic OH. An allyl ester and allyl ether group were installed onto TCF-OH and DCI-OH, constructing four potential CO fluorescent probes, TCF-ester, TCF-ether, DCI-ester, and DCI-ether. Our data revealed that ester bonds of TCF-ester and DCI-ester could completely hydrolyze in 20 min, but ether bonds in TCF-ether and DCI-ether tolerate the hydrolysis of BSA and no released fluorescence was observed even up to 2 h. Moreover, passing through the screen, it was concluded that TCF-ether is superior to DCI-ether due to its higher reactivity in a Pd0-mediated Tsuji-Trost reaction. Also, the large stokes shift of TCF-OH, absorption and emission at 408 nm and 618 nm respectively, make TCF-ether desirable for fluorescent imaging because of differentiating signals from the excitation light source. Lastly, TCF-ether has been successfully applied to the detection of CO in H9C2 cells.

The importance of and difficulties involved in creating molecular probes for a carbon monoxide gasotransmitter

“…One can ponder over whether the palladium-catalyzed process in the Tsuji-Trost probe system may be affected by the reaction conditions. 134 The reducing capability of CO to effectuate the transformation of Pd 2+ into Pd 0 , subsequently enabling Pd 0 to catalyse the de-allylation Tsuji-Trost reaction, might confront a substantial hurdle presented by the manifold reductive substances possessing the ability to revert Pd 2+ to Pd 0 . While this reality remains to be extensively examined, its apparent nature calls for additional comprehensive scrutiny by researchers.…”

“…One can ponder over whether the palladium-catalyzed process in the Tsuji–Trost probe system may be affected by the reaction conditions. 134…”