Postgraduate researcher Johannes Woehrstein, Wyss Institute for Biologically Inspired Engineering, Center for Life Science, Boston, Massachusetts, United States

Pushing the Limits in Fluorescence Microscopy: Highly Multiplexed Super-Resolution Imaging With DNA-PAINT

Super-resolution fluorescence microscopy is a powerful tool for biological research, but obtaining multiplexed images for a large number of distinct target species remains challenging. In DNA-PAINT the transient binding of short fluorescently labeled oligonucleotides is used for simple and easy-to-implement multiplexed super-resolution imaging that achieves sub-10-nm spatial resolution. By labeling target proteins with antibody-DNA conjugates, DNA-PAINT is also applicable to in situ cell imaging in two and three dimensions.

We recently used the unique programmability of DNA binding interactions in DNA-PAINT imaging to develop a novel multiplexing approach we call Exchange-PAINT. This allows sequential imaging of multiple targets using only a single dye and a single laser source. Using Exchange-PAINT, we experimentally demonstrated ten-color sub-10 nm super-resolution imaging in vitro as well as four-color two-dimensional imaging and three-color 3D imaging of proteins in fixed cells.