Programming in situ immunofluorescence intensities through interchangeable reactions of dynamic DNA complexes
Schweller, Ryan M.
Duose, Dzifa Y.
Hittelman, Walter N.
Diehl, Michael R.
The regulation of antibody reporting intensities is critical to various in situ fluorescence imaging analyses. While such control is often necessary to visualize sparse molecular targets, the ability to tune marker intensities is also essential for highly multiplexed imaging strategies where marker reporting levels must be tuned to both optimize dynamic detection ranges and minimize crosstalk between different signals. Existing chemical amplification approaches generally lack such control. Here, we demonstrate that linear and branched DNA complexes can be designed to function as interchangeable building blocks that can be assembled into organized, fluorescence reporting complexes. We show that the ability to program DNA strand displacement reactions between these complexes offer new opportunities to deterministically tune the number of dyes that are coupled to individual antibodies in order to both increase and controllably balance marker levels within fixed cells.
antibodies; immunofluorescence; DNA strand displacement; dynamic DNA; multiplexed detection