Journal
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
Volume 143, Issue 41, Pages 16976-16992Publisher
AMER CHEMICAL SOC
DOI: 10.1021/jacs.1c06094
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Funding
- Natural Sciences and Engineering Research Council of Canada (NSERC)
- Canada Foundation for Innovation
- British Columbia Knowledge Development Fund
- University of British Columbia (UBC)
- NSERC for a Canada Graduate Scholarship
- NSERC CREATE NanoMat program
- Canada First Research Excellence Fund
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In this study, deep blue fluorophores were designed to improve resistance to photobleaching, two-photon absorption cross sections, and fluorescence quantum yields using structural constraints. The resulting water-soluble Pdots exhibited high brightness, improved signal-to-noise ratio, and resistance to photobleaching, making them suitable for single-particle visualization and biological imaging applications such as immunolabeling of breast cancer cells.
Semiconducting polymer dots (Pdots) have emerged as versatile probes for bioanalysis and imaging at the singleparticle level. Despite their utility in multiplexed analysis, deep blue Pdots remain rare due to their need for high-energy excitation and sensitivity to photobleaching. Here, we describe the design of deep blue fluorophores using structural constraints to improve resistance to photobleaching, two-photon absorption cross sections, and fluorescence quantum yields using the hexamethylazatriangulene motif. Scanning tunneling microscopy was used to characterize the electronic structure of these chromophores on the atomic scale as well as their intrinsic stability. The most promising fluorophore was functionalized with a polymerizable acrylate handle and used to give deep-blue fluorescent acrylic polymers with M-n > 18 kDa and D < 1.2. Nanoprecipitation with amphiphilic polystyrene-graf t-(carboxylate-terminated poly(ethylene glycol)) gave water-soluble Pdots with blue fluorescence, quantum yields of 0.81, and molar absorption coefficients of (4 +/- 2) x 10(8) M-1 cm(-1). This high brightness facilitated single-particle visualization with dramatically improved signal-to-noise ratio and photobleaching resistance versus an unencapsulated dye. The Pdots were then conjugated with antibodies for immunolabeling of SK-BR3 human breast cancer cells, which were imaged using deep blue fluorescence in both one- and two-photon excitation modes.
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