DNA-points accumulation for imaging in nanoscale topography (DNA-PAINT) has emerged as a powerful super-resolution imaging technique capable of visualizing biological and synthetic structures with nanometer precision. By exploiting the transient binding of short fluorescently labeled DNA “imager” strands to complementary “docking” strands attached to target molecules, DNA-PAINT enables multiplexed visualization of targets with molecular resolution. We expand the existing repertoire of speed-optimized DNA sequences for DNA-PAINT imaging to drive visualization of up to ten different targets in a sequential manner with molecular resolution. Our speed-optimized imager-docking strand pairs minimize crosstalk and maximize hybridization rates, enabling rapid image acquisition without compromising spatial resolution. We demonstrate this ability by visualizing diverse nuclear targets in single cells and subsequently characterizing their relative spatial distribution. We further outline the ability of the technique to visualize and detect nanoscale reorganization in the chromatin landscape upon transcription inhibition. The ability to image as many as ten targets using this strategy at accelerated speed will drive its further adoption for diverse cellular imaging applications.
Anand et al. (Sun,) studied this question.