title: Super-Resolution Imaging and Quantitative Analysis of Membrane Protein/Lipid Raft Clustering Mediated by Cell-Surface Self-Assembly of Hybrid Nanoconjugates.
abstract: Super-resolution imaging was used to quantify organizational changes in the plasma membrane after treatment with hybrid nanoconjugates. The nanoconjugates crosslinked CD20 on the surface of malignant B cells, thereby inducing apoptosis. Super-resolution images were analyzed by using pair-correlation analysis to determine cluster size and to count the average number of molecules in the clusters. The role of lipid rafts was investigated by pre-treating cells with a cholesterol chelator and actin destabilizer to prevent lipid raft formation. Lipid raft cluster size correlated with apoptosis induction after treatment with the nanoconjugates. Lipid raft clusters had radii of ∼ 200 nm in cells treated with the hybrid nanoconjugates. Super-resolution images provided precise molecule location coordinates that could be used to determine density of bound conjugates, cluster size, and number of molecules per cluster.
authors.affiliation: Department of Bioengineering, University of Utah, 20 S. 2030 E., Room 108, Salt Lake City, UT 84112 (USA).
authors.forename: Jonathan M
authors.initials: JM
authors.lastname: Hartley
authors.affiliation: Department of Pharmaceutics and Pharmaceutical Chemistry, University of Utah, 30 S. 2000 E Room 301, Salt Lake City, UT 84112 (USA).
authors.forename: Te-Wei
authors.initials: TW
authors.lastname: Chu
authors.affiliation: Department of Chemistry, University of Utah, 315 S. 1400 E, Room 2020, Salt Lake City, UT 84112 (USA).
authors.forename: Eric M
authors.initials: EM
authors.lastname: Peterson
authors.affiliation: Department of Pharmaceutics and Pharmaceutical Chemistry, University of Utah, 30 S. 2000 E Room 301, Salt Lake City, UT 84112 (USA).
authors.forename: Rui
authors.initials: R
authors.lastname: Zhang
authors.affiliation: Department of Pharmaceutics and Pharmaceutical Chemistry, University of Utah, 30 S. 2000 E Room 301, Salt Lake City, UT 84112 (USA).
authors.forename: Jiyuan
authors.initials: J
authors.lastname: Yang
authors.affiliation: Department of Chemistry, University of Utah, 315 S. 1400 E, Room 2020, Salt Lake City, UT 84112 (USA).
authors.forename: Joel
authors.initials: J
authors.lastname: Harris
authors.affiliation: Department of Bioengineering, University of Utah, 20 S. 2030 E., Room 108, Salt Lake City, UT 84112 (USA). jindrich.kopecek@utah.edu.|Department of Pharmaceutics and Pharmaceutical Chemistry, University of Utah, 30 S. 2000 E Room 301, Salt Lake City, UT 84112 (USA). jindrich.kopecek@utah.edu.
authors.forename: Jindřich
authors.initials: J
authors.lastname: Kopeček
mesh_terms: D017209:Apoptosis; D002462:Cell Membrane / Q000378:metabolism; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism*; D008954:Models, Biological; D057054:Molecular Imaging; D058726:Nanoconjugates / Q000737:chemistry*
chemical_list: D008565:Membrane Proteins; D058726:Nanoconjugates
keywords: dSTORM; nanomedicine; nanotechnology; self-assembly