Integrated Micropillar Polydimethylsiloxane Accurate CRISPR Detection (IMPACT) System for Rapid Viral DNA Sensing

Kenneth N. Hass; Mengdi Bao; Qian He; Myeongkee Park; Peiwu Qin; Ke Du

The extended surface provided by high-aspect ratio micropillars significantly increases the reporter probe binding capacity. To demonstrate this, we compared the number of captured DNA molecules on the micropillar channel with the flat channel (Fig. 4) . With an input of 1x10 4 nmoles, the retrieved DNA from the micropillar channel has an integrated intensity of ~2,000 counts, which is much higher than the flat surface (~900 counts). With an input of 1x10 5 nmoles, the retrieved DNA from the micropillar channel has an integrated intensity of ~5,500 counts and is ~40% higher than the flat channel. With an input of 1x10 6 nmoles, the micropillar channel still shows higher signal than the flat channel but is comparable to 1x10 5 nmoles sample, indicating that the channel is saturated with an input of ~1x10 -5 nmoles. Thus, we used 1x10 5 nmoles reporter probes in our CRISPR detection as higher load could cause unnecessary background.