A ratiometric nanoprobe for biosensing based on green fluorescent graphitic carbon nitride nanosheets as an internal reference and quenching platform

Abstract The integrating of high fluorescent quenching capability of two-dimensional nanomaterials with dye-labelled ssDNA as nanoprobes has attracted increasing interest for biosensing applications. However, this absolute intensity-dependent single fluorescent signal may be influenced by target concentration-independent factors. To overcome this challenge, a ratiometric nanoprobe was developed by utilizing green fluorescent phenyl-doped carbon nitride (PDCN) nanosheets as an internal reference and quenching platform. 5-carboxy-X-rhodamine-labelled anti-adenosine aptamer was used as a signal probe. PDCN nanosheets quenched the fluorescence of the absorbed signal probe while kept their own fluorescence constantly. Upon addition of adenosine, the formation of adenosine-aptamer complexes led to desorption of the signal probe from the surface of PDCN nanosheets, resulted in the fluorescent recovery of the signal probe. The ratio of the fluorescent enhancement of the signal probe to the inherent fluorescence of PDCN nanosheets was used to quantitatively measure adenosine. The limit of detection for adenosine was 6.86 μM. Finally, the ratiometric nanoprobe was applied to determine adenosine in serum samples.