Quantitative photo activated localization microscopy: unraveling the effects of photoblinking

PLoS One. 2011;6(7):e22678. doi: 10.1371/journal.pone.0022678. Epub 2011 Jul 26.


In this work we discuss how to use photophysical information for improved quantitative measurements using Photo Activated Localization Microscopy (PALM) imaging. We introduce a method that reliably estimates the number of photoblinking molecules present in a biological sample and gives a robust way to quantify proteins at the single-cell level from PALM images. We apply this method to determine the amount of β2 adrenergic receptor, a prototypical G Protein Coupled Receptor, expressed on the plasma membrane of HeLa cells.

MeSH terms

  • Computer Simulation
  • HeLa Cells
  • Humans
  • Light*
  • Luminescent Proteins / metabolism
  • Microscopy / methods*
  • Receptors, Adrenergic, beta-2 / metabolism


  • Luminescent Proteins
  • Receptors, Adrenergic, beta-2