Fluorescent labels have become ubiquitous in biology, staining cell membranes for imaging, proteins of interest for tracking, and intracellular components for quantification. Furthermore, the use of fluorescent tagged secondary antibodies in immunofluorescence protocols has allowed for higher specificity targeting and localization of these labels. The Armitage lab seeks to enhance this growing field through further development of brightly fluorescent bottlebrush polymer (BBP) nanotags, functionalized with dsDNA brush "bristles" loaded with co-bisintercalating FRET dye pairs. A previous BBP nanotag with intercalated donor dyes and acceptor dyes covalently linked to the bristle tips has shown enhanced energy transfer resulting in fluorescent levels above comparable commercial counterparts. Switching to a co-bisintercalating dye pair has eliminated a synthetic step, while also introducing additional ease of tunability in sensor brightness and fluorescent range. Synthesis of these BBP scaffolds is controlled through atom transfer radical polymerization (ATRP), while the tags are assembled using psuedoligandless Cu(I) catalyzed click reactions. We demonstrate the loading of these BBP nanotags with optimized ratios of different bisintercalating FRET dye pairs, characterized via fluorescence spectroscopy. In addition, when conjugated to antibody or aptamer affinity reagents, these probes can be used to detect molecular targets in flow cytometry.

Co-bisintercalating FRET-based fluorescent bottlebrush polymer nanotags

253rd National Meeting (2017)

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