2. Academic Validation
  3. DNA-Origami-Based Fluorescence Brightness Standards for Convenient and Fast Protein Counting in Live Cells

DNA-Origami-Based Fluorescence Brightness Standards for Convenient and Fast Protein Counting in Live Cells

  • Nano Lett. 2020 Dec 9;20(12):8890-8896. doi: 10.1021/acs.nanolett.0c03925.
Nathan D Williams 1 2,  Ane Landajuela 2 3,  Ravi Kiran Kasula 1,  Wenjiao Zhou 1 2,  John T Powell 1 2,  Zhiqun Xi 1,  Farren J Isaacs 4 5 6,  Julien Berro 1 2 7,  Derek Toomre 1,  Erdem Karatekin 2 3 7 8,  Chenxiang Lin 1 2
Affiliations

Affiliations

  • 1 Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut 06520, United States.
  • 2 Nanobiology Institute, Yale University, West Haven, Connecticut 06516, United States.
  • 3 Department of Cellular and Molecular Physiology, Yale University School of Medicine, New Haven, Connecticut 06520, United States.
  • 4 Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, Connecticut 06520, United States.
  • 5 Department of Biomedical Engineering, Yale University, New Haven, Connecticut 06520, United States.
  • 6 Systems Biology Institute, Yale University, West Haven, Connecticut 06516, United States.
  • 7 Department of Molecular Biophysics and Biochemistry, New Haven, Connecticut 06520, United States.
  • 8 SPPIN - Saints-Pères Paris Institute for the Neurosciences, Centre National de la Recherche Scientifique (CNRS), Université de Paris, F-75006 Paris, France.
PMID: 33164530 DOI: 10.1021/acs.nanolett.0c03925
Abstract

Fluorescence microscopy has been one of the most discovery-rich methods in biology. In the digital age, the discipline is becoming increasingly quantitative. Virtually all biological laboratories have access to fluorescence microscopes, but abilities to quantify biomolecule copy numbers are limited by the complexity and sophistication associated with current quantification methods. Here, we present DNA-origami-based fluorescence brightness standards for counting 5-300 copies of proteins in Bacterial and mammalian cells, tagged with fluorescent proteins or membrane-permeable organic dyes. Compared to conventional quantification techniques, our brightness standards are robust, straightforward to use, and compatible with nearly all fluorescence imaging applications, thereby providing a practical and versatile tool to quantify biomolecules via fluorescence microscopy.

Keywords

Bioconjugation; Brightness standard; DNA origami; Fluorescent protein; Live cell imaging; Quantitative microscopy.

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