Single-molecule Super-resolution Imaging of Phosphatidylinositol 4,5-bisphosphate in the Plasma Membrane with Novel Fluorescent Probes. J Vis Exp 2016 Oct 15(116)
Date
11/03/2016Pubmed ID
27805608Pubmed Central ID
PMC5092206DOI
10.3791/54466Scopus ID
2-s2.0-84992505061 (requires institutional sign-in at Scopus site) 7 CitationsAbstract
Phosphoinositides in the cell membrane are signaling lipids with multiple cellular functions. Phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2) is a determinant phosphoinositide of the plasma membrane (PM), and it is required to modulate ion channels, actin dynamics, exocytosis, endocytosis, intracellular signaling, and many other cellular processes. However, the spatial organization of PI(4,5)P2 in the PM is controversial, and its nanoscale distribution is poorly understood due to the technical limitations of research approaches. Here by utilizing single molecule localization microscopy and the Pleckstrin Homology (PH) domain based dual color fluorescent probes, we describe a novel method to visualize the nanoscale distribution of PI(4,5)P2 in the PM in fixed membrane sheets as well as live cells.
Author List
Ji C, Lou XAuthor
Xuelin Lou PhD Professor in the Cell Biology, Neurobiology and Anatomy department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
ActinsCell Membrane
Fluorescent Dyes
Phosphatidylinositol 4,5-Diphosphate
Phosphatidylinositol Phosphates
Phosphatidylinositols
Single Molecule Imaging
