Unit

UNIT 4.21 Photoactivated Localization Microscopy (PALM) of Adhesion Complexes

  1. Hari Shroff,
  2. Helen White,
  3. Eric Betzig

Published Online: 1 DEC 2008

DOI: 10.1002/0471143030.cb0421s41

Current Protocols in Cell Biology

Current Protocols in Cell Biology

How to Cite

Shroff, H., White, H. and Betzig, E. 2008. Photoactivated Localization Microscopy (PALM) of Adhesion Complexes. Current Protocols in Cell Biology. 41:4.21:4.21.1–4.21.27.

Author Information

  1. Howard Hughes Medical Institute, Janelia Farm Research Campus, Ashburn, Virginia

Publication History

  1. Published Online: 1 DEC 2008
  2. Published Print: DEC 2008

This is not the most recent version of the article. View current version (1 MAR 2013)

Abstract

Key to understanding a protein's biological function is the accurate determination of its spatial distribution inside a cell. Although fluorescent protein markers allow the targeting of specific proteins with molecular precision, much of this information is lost when the resultant fusion proteins are imaged with conventional, diffraction-limited optics. In response, several imaging modalities that are capable of resolution below the diffraction limit (∼200 nm) have emerged. Here, both single- and dual-color superresolution imaging of biological structures using photoactivated localization microscopy (PALM) are described. The examples discussed focus on adhesion complexes: dense, protein-filled assemblies that form at the interface between cells and their substrata. A particular emphasis is placed on the instrumentation and photoactivatable fluorescent protein (PA-FP) tags necessary to achieve PALM images at ∼20 nm resolution in 5 to 30 min in fixed cells. Curr. Protoc. Cell Biol. 41:4.21.1-4.21.27. © 2008 by John Wiley & Sons, Inc.

Keywords:

  • PALM;
  • superresolution;
  • adhesion complex;
  • fluorescent proteins