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Quantitative Photo Activated Localization Microscopy: Unraveling the Effects of Photoblinking

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  • Paolo Annibale
  • Stefano Vanni
  • Marco Scarselli
  • Ursula Rothlisberger
  • Aleksandra Radenovic

Abstract

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.

Suggested Citation

  • Paolo Annibale & Stefano Vanni & Marco Scarselli & Ursula Rothlisberger & Aleksandra Radenovic, 2011. "Quantitative Photo Activated Localization Microscopy: Unraveling the Effects of Photoblinking," PLOS ONE, Public Library of Science, vol. 6(7), pages 1-8, July.
    • Handle: RePEc:plo:pone00:0022678
    DOI: 10.1371/journal.pone.0022678
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    References listed on IDEAS

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    1. Long Cai & Nir Friedman & X. Sunney Xie, 2006. "Stochastic protein expression in individual cells at the single molecule level," Nature, Nature, vol. 440(7082), pages 358-362, March.
    2. Robert M. Dickson & Andrew B. Cubitt & Roger Y. Tsien & W. E. Moerner, 1997. "On/off blinking and switching behaviour of single molecules of green fluorescent protein," Nature, Nature, vol. 388(6640), pages 355-358, July.
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    Cited by:

    1. Carla Coltharp & Rene P Kessler & Jie Xiao, 2012. "Accurate Construction of Photoactivated Localization Microscopy (PALM) Images for Quantitative Measurements," PLOS ONE, Public Library of Science, vol. 7(12), pages 1-15, December.
    2. Aske L. Ejdrup & Matthew D. Lycas & Niels Lorenzen & Ainoa Konomi & Freja Herborg & Kenneth L. Madsen & Ulrik Gether, 2022. "A density-based enrichment measure for assessing colocalization in single-molecule localization microscopy data," Nature Communications, Nature, vol. 13(1), pages 1-10, December.

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