Quantitative Detection and Imaging of Multiple Biological Molecules in Living Cells for Cell Screening

Because of insufficient information, a single biomarker is not sufficient for early diagnosis of cancer, whereas sensitive and selective detection of multiple biomolecules can significantly reduce analysis time, sample size, and accurately perform cell screening in early cancer. Therefore, the development of a noninvasive strategy that can simultaneously quantify multiple biomarkers (i.e., nucleic acids, proteins, and small molecules) in a single cell is particularly important. Herein, a universal sensing system (functional DNA@mesoporous silica nanoparticles (MSN)-Black Hole Quencher-rhodamine 6G (RhB), FDSBR), which is based on the combination of functionalized DNA and smart responsive nanomaterial, was successfully constructed. After incubation with the cells, different types of targets trigger the strand displacement reaction to release the fluorophore-labeled nucleic acids as the output signals to reflect the intracellular level of the telomerase and adenosine triphosphate (ATP), respectively. Simultaneously, intracellular miR-21 can be clearly indicated by the restored fluorescence of RhB when the caged double-stranded DNA was substituted into single-stranded DNA to open the pore. The concentrations of intracellular telomerase, miR-21, and ATP were identified successfully in three cell lines at the single-cell level. The results show that the contents of three biomolecules have significant differences in the three model cell lines and provide a promising route for developing innovative early disease diagnosis and cell screening assay.