Open AccessDNA-Origami-Based Fluorescence Brightness Standards for Convenient and Fast Protein Counting in Live Cells
Author(s) -
Nathan D. Williams,
Ane Landajuela,
Ravi Kiran Kasula,
Wanting Zhou,
John T. Powell,
Zhiqun Xi,
Farren J. Isaacs,
Julien Berro,
Derek Toomre,
Erdem Karatekin,
Chenxiang Lin
Publication year - 2020
Publication title -
nano letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 4.853
H-Index - 488
eISSN - 1530-6992
pISSN - 1530-6984
DOI - 10.1021/acs.nanolett.0c03925
Subject(s) - fluorescence , biomolecule , fluorescence microscope , brightness , microscope , microscopy , nanotechnology , dna origami , biophysics , materials science , chemistry , biological system , biology , optics , physics , nanostructure
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.