Quantitative dynamic footprinting microscopy reveals mechanisms of neutrophil rolling

Nat Methods. 2010 Oct;7(10):821-4. doi: 10.1038/nmeth.1508. Epub 2010 Sep 26.

Abstract

We introduce quantitative dynamic footprinting microscopy to resolve neutrophil rolling on P-selectin. We observed that the footprint of a rolling neutrophil was fourfold larger than previously thought, and that P-selectin-PSGL-1 bonds were relaxed at the leading edge of the rolling cell, compressed under the cell center, and stretched at the trailing edge. Each rolling neutrophil formed three to four long tethers that extended up to 16 μm behind the rolling cell.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Green Fluorescent Proteins / genetics
  • Leukocyte Rolling / physiology*
  • Membrane Glycoproteins / genetics*
  • Mice
  • Mice, Transgenic
  • Microfluidic Analytical Techniques / instrumentation
  • Microfluidic Analytical Techniques / methods
  • Microscopy, Fluorescence / instrumentation
  • Microscopy, Fluorescence / methods*
  • Muramidase / genetics
  • Neutrophils / metabolism
  • Neutrophils / physiology*
  • Neutrophils / ultrastructure
  • P-Selectin / genetics*
  • Protein Footprinting / instrumentation
  • Protein Footprinting / methods*

Substances

  • Membrane Glycoproteins
  • P-Selectin
  • P-selectin ligand protein
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • Muramidase
  • lysozyme M, mouse