FALCON: fast and unbiased reconstruction of high-density super-resolution microscopy data

Sci Rep. 2014 Apr 3:4:4577. doi: 10.1038/srep04577.

Abstract

Super resolution microscopy such as STORM and (F)PALM is now a well known method for biological studies at the nanometer scale. However, conventional imaging schemes based on sparse activation of photo-switchable fluorescent probes have inherently slow temporal resolution which is a serious limitation when investigating live-cell dynamics. Here, we present an algorithm for high-density super-resolution microscopy which combines a sparsity-promoting formulation with a Taylor series approximation of the PSF. Our algorithm is designed to provide unbiased localization on continuous space and high recall rates for high-density imaging, and to have orders-of-magnitude shorter run times compared to previous high-density algorithms. We validated our algorithm on both simulated and experimental data, and demonstrated live-cell imaging with temporal resolution of 2.5 seconds by recovering fast ER dynamics.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Algorithms*
  • Animals
  • COS Cells
  • Carbocyanines / chemistry
  • Cell Line
  • Chlorocebus aethiops
  • Endoplasmic Reticulum / metabolism
  • Fluorescent Dyes / chemistry
  • Humans
  • Microscopy, Fluorescence*
  • Microtubules / chemistry
  • Microtubules / metabolism
  • Signal-To-Noise Ratio

Substances

  • Alexa Fluor 647
  • Carbocyanines
  • Fluorescent Dyes