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Sci Adv. 2017 Jun 21;3(6):e1602128. doi: 10.1126/sciadv.1602128. eCollection 2017 Jun.

Sub-100-nm metafluorophores with digitally tunable optical properties self-assembled from DNA.

Woehrstein JB1,2,3, Strauss MT1,2,3, Ong LL1,4, Wei B1,5, Zhang DY1,5, Jungmann R1,2,3,5, Yin P1,5.

Author information

1
Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA 02115, USA.
2
Department of Physics and Center for NanoScience, Ludwig Maximilian University, 80539 Munich, Germany.
3
Max Planck Institute of Biochemistry, 82152 Martinsried near Munich, Germany.
4
Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
5
Department of Systems Biology, Harvard Medical School, Boston, MA 02115, USA.

Abstract

Fluorescence microscopy allows specific target detection down to the level of single molecules and has become an enabling tool in biological research. To transduce the biological information to an imageable signal, we have developed a variety of fluorescent probes, such as organic dyes or fluorescent proteins with different colors. Despite their success, a limitation on constructing small fluorescent probes is the lack of a general framework to achieve precise and programmable control of critical optical properties, such as color and brightness. To address this challenge, we introduce metafluorophores, which are constructed as DNA nanostructure-based fluorescent probes with digitally tunable optical properties. Each metafluorophore is composed of multiple organic fluorophores, organized in a spatially controlled fashion in a compact sub-100-nm architecture using a DNA nanostructure scaffold. Using DNA origami with a size of 90 × 60 nm2, substantially smaller than the optical diffraction limit, we constructed small fluorescent probes with digitally tunable brightness, color, and photostability and demonstrated a palette of 124 virtual colors. Using these probes as fluorescent barcodes, we implemented an assay for multiplexed quantification of nucleic acids. Additionally, we demonstrated the triggered in situ self-assembly of fluorescent DNA nanostructures with prescribed brightness upon initial hybridization to a nucleic acid target.

KEYWORDS:

Biophysics; DNA nanotechnology; Metafluorophores; Multiplexed Imaging; fluorescence microscopy; self assembly; single molecule fluorescence

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