A rhodol-derived probe for intracellular biothiols imaging and rapid labelling of sulfhydryl-containing proteins

Biological thiols are crucial in biological processes. Therefore, a probe capable of targeting and imaging biothiols in living cells, as well as rapidly labelling the thiols in proteins, is desirable and imperative. Herein, we designed and synthesized FRB-T containing α, β-unsaturated ketones to recognize thiols for detecting biothiols and selectively labelling the sulfhydryl-containing proteins. The probe initially exhibited weak fluorescence and then performed 1, 4-Michael addition reaction between its α, β-unsaturated ketone and thiols, along with the spiro ring-opening, leading to remarkable fluorescence enhancement and colour changes. The specificity of FRB-T towards thiols was high, and its detection limits were 70 nM for cysteine (Cys) and 1.51 μg/mL for ovalbumin (OVA). Furthermore, FRB-T has also been successfully applied to biothiols imaging in Hela cells, and rapid labelling of the sulfhydryl-containing proteins without washing step.