A practical approach for the detection of DNA nanostructures in single live human cells by fluorescence microscopy

Methods. 2014 May 15;67(2):185-92. doi: 10.1016/j.ymeth.2014.01.009. Epub 2014 Jan 17.

Abstract

In the last decade, in vivo studies have revealed that even subtle differences in size, concentration of components, cell cycle stage, make the cells in a population respond differently to the same stimulus. In order to characterize such complexity of behavior and shed more light on the functioning and communication amongst cells, researchers are developing strategies to study single live cells in a population. In this paper, we describe the methods to design and prepare DNA-based fluorescent tetrahedral nanostructures, to deliver them to live cells and characterize such cells with epifluorescence microscopy. We report that HeLa cells internalize these nanostructures spontaneously with a higher efficiency with respect to single-stranded or double-stranded oligonucleotides. Our findings suggest that DNA tetrahedra could serve as a platform for the realization of a series of multifunctional intracellular biosensors for the analysis of single live cells.

Keywords: DNA nanotechnology; Flow cytofluorimetry; Fluorescence microscopy; Live-cell imaging; Self-assembled nanostructures.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA / chemistry*
  • DNA / ultrastructure
  • Fluorescent Dyes / chemistry
  • HeLa Cells
  • Humans
  • Microscopy, Fluorescence
  • Nanostructures / chemistry
  • Nanostructures / ultrastructure
  • Nucleic Acid Conformation
  • Oligonucleotides / chemistry*

Substances

  • Fluorescent Dyes
  • Oligonucleotides
  • DNA