Real-time analysis and visualization for single-molecule based super-resolution microscopy

PLoS One. 2013 Apr 30;8(4):e62918. doi: 10.1371/journal.pone.0062918. Print 2013.

Abstract

Accurate multidimensional localization of isolated fluorescent emitters is a time consuming process in single-molecule based super-resolution microscopy. We demonstrate a functional method for real-time reconstruction with automatic feedback control, without compromising the localization accuracy. Compatible with high frame rates of EM-CCD cameras, it relies on a wavelet segmentation algorithm, together with a mix of CPU/GPU implementation. A combination with Gaussian fitting allows direct access to 3D localization. Automatic feedback control ensures optimal molecule density throughout the acquisition process. With this method, we significantly improve the efficiency and feasibility of localization-based super-resolution microscopy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Algorithms
  • Animals
  • COS Cells
  • Chlorocebus aethiops
  • Image Processing, Computer-Assisted / instrumentation
  • Image Processing, Computer-Assisted / methods
  • Microscopy, Fluorescence / instrumentation
  • Microscopy, Fluorescence / methods*
  • Software

Grants and funding

This study was supported by CNRS; ANR; Regional Council of Aquitaine; Fondation pour la Recherche Médicale; European Union’s seventh framework program for research and development for the Marie-Curie-Intra European Fellowship; ERC; ERA-NET; German Ministry of Research and Education. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.