Journal
CHEMBIOCHEM
Volume 13, Issue 18, Pages 2722-2728Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/cbic.201200525
Keywords
antibodies; DNA-strand displacement; dynamic DNA; immunofluorescence; multiplexed detection
Funding
- National Institutes of Health [1R21A147912]
- Welch Foundation [C-1625]
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The regulation of antibody reporting intensities is critical to various in situ fluorescence-imaging analyses. Although such control is often necessary to visualize sparse molecular targets, the ability to tune marker intensities is also essential for highly multiplexed imaging strategies in which marker reporting levels must be tuned both to optimize dynamic detection ranges and to 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 offers new opportunities to deterministically tune the number of dyes that are coupled to individual antibodies in order both to increase and controllably balance marker reporting levels within fixed cells.
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