Non-amplified quantitative detection of nucleic acid sequences using a gold nanoparticle probe set and field-emission scanning electron microscopy

For the precise detection of the number of expressed biomarkers at the single-cell level, we have developed a method of quantifying and specifying target DNA fragments by using a set of gold nanoparticles as labels and field-emission scanning electron microscopy (FE-SEM) to measure the number and sizes of gold nanoparticles attached to target samples. One or more target DNAs on a substrate were labeled with a set of different-sized gold nanoparticle probes having complementary sequences to different target candidates. The type and number of the target DNAs having a specific sequence were identified by counting the attached nanoparticles of a specific size in FE-SEM images. The results evaluated using a DNA microarray showed high specificity and sensitivity, and a linear correlation between the number of attached particles and the target DNA concentration, indicating the feasibility of quantitative detection in the femtomolar to nanomolar concentration range.