Three-Dimensional Tracking of Quantum Dot-Conjugated Molecules in Living Cells

Methods Mol Biol. 2018:1814:425-448. doi: 10.1007/978-1-4939-8591-3_26.

Abstract

Here, we describe protocols for three-dimensional tracking of single quantum dot-conjugated molecules with nanometer accuracy in living cells using conventional fluorescence microscopy. The technique exploits out-of-focus images of single emitters combined with an automated pattern-recognition open-source software that fits the images with proper model functions to extract the emitter coordinates. We describe protocols for targeting quantum dots to both membrane components and cytosolic proteins.

Keywords: Cytosolic proteins; Lipids; Living cells; Out-of-focus imaging; Plasma membrane proteins; Quantum dots; Single-molecule localization; Three-dimensional tracking.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Algorithms
  • Calibration
  • Cell Line, Tumor
  • Cell Survival
  • Cytosol / metabolism
  • Data Analysis
  • G(M1) Ganglioside / metabolism
  • Humans
  • Image Processing, Computer-Assisted
  • Imaging, Three-Dimensional*
  • Membrane Proteins / metabolism
  • Microscopy, Fluorescence / methods*
  • Quantum Dots / chemistry*
  • Staining and Labeling

Substances

  • Membrane Proteins
  • G(M1) Ganglioside