title: Super-resolution Analysis of TCR-Dependent Signaling: Single-Molecule Localization Microscopy.
abstract: Single-molecule localization microscopy (SMLM) comprises methods that produce super-resolution images from molecular locations of single molecules. These techniques mathematically determine the center of a diffraction-limited spot produced by a fluorescent molecule, which represents the most likely location of the molecule. Only a small cohort of well-separated molecules is visualized in a single image, and then many images are obtained from a single sample. The localizations from all the images are combined to produce a super-resolution picture of the sample. Here we describe the application of two methods, photoactivation localization microscopy (PALM) and direct stochastic optical reconstruction microscopy (dSTORM), to the study of signaling microclusters in T cells.
authors.affiliation: Laboratory of Cellular and Molecular Biology, Center for Cancer Research, National Cancer Institute, Bethesda, MD, 20892-4256, USA.
authors.forename: Valarie A
authors.initials: VA
authors.lastname: Barr
authors.affiliation: Laboratory of Cellular and Molecular Biology, Center for Cancer Research, National Cancer Institute, Bethesda, MD, 20892-4256, USA.
authors.forename: Jason
authors.initials: J
authors.lastname: Yi
authors.affiliation: Laboratory of Cellular and Molecular Biology, Center for Cancer Research, National Cancer Institute, Bethesda, MD, 20892-4256, USA. samelsonl@helix.nih.gov.
authors.forename: Lawrence E
authors.initials: LE
authors.lastname: Samelson
mesh_terms: D006801:Humans; D019169:Jurkat Cells; D057054:Molecular Imaging / Q000379:methods*; D011948:Receptors, Antigen, T-Cell / Q000276:immunology*; D015398:Signal Transduction / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology*
chemical_list: D011948:Receptors, Antigen, T-Cell
keywords: Direct stochastic optical resolution microscopy; Microclusters; Photoactivation localization microscopy; Single-molecule localization microscopy; Super-resolution microscopy; T cell