Nature Methods
- 5, 155 - 157 (2008)
Published online: 13 January 2008; | doi:10.1038/nmeth.1176
High-density mapping of single-molecule trajectories with photoactivated localization microscopySuliana Manley1, Jennifer M Gillette1, George H Patterson1, Hari Shroff2, Harald F Hess2, Eric Betzig2 & Jennifer Lippincott-Schwartz11
National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892, USA. 2
Howard Hughes Medical Institute, Janelia Farm Research Campus, Ashburn, Virginia 20147, USA.
Correspondence should be addressed to Jennifer Lippincott-Schwartz jlippin@helix.nih.gov We combined photoactivated localization microscopy (PALM) with live-cell single-particle tracking to create a new method termed sptPALM. We created spatially resolved maps of single-molecule motions by imaging the membrane proteins Gag and VSVG, and obtained several orders of magnitude more trajectories per cell than traditional single-particle tracking enables. By probing distinct subsets of molecules, sptPALM can provide insight into the origins of spatial and temporal heterogeneities in membranes.
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