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Programming in Situ Immunofluorescence Intensities through Interchangeable Reactions of Dynamic DNA Complexes
Author(s) -
Zimak Jan,
Schweller Ryan M.,
Duose Dzifa Y.,
Hittelman Walter N.,
Diehl Michael R.
Publication year - 2012
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201200525
Subject(s) - in situ , dna , fluorescence , computer science , multiplexing , immunofluorescence , biophysics , biological system , chemistry , nanotechnology , computational biology , materials science , combinatorial chemistry , biology , antibody , physics , optics , biochemistry , genetics , telecommunications , organic chemistry
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.