title: Viewing dynamic interactions of proteins and a model lipid membrane with atomic force microscopy.
abstract: The information covered in this chapter will present a model homogenous membrane preparation technique and dynamic imaging procedure that can be successfully applied to more than one type of lipid study and atomic force microscope (AFM) instrument setup. The basic procedural steps have been used with an Asylum Research MFP-3D BIO and the Bruker (formerly, Veeco) BioScope. The AFM imaging protocol has been supplemented by procedures (not to be presented in this chapter) of ellipsometry, standardized western blotting, and dot-blots to verify appropriate purity and activity of all experimental molecular components; excellent purity and activity level of the lipids, proteins, and drug(s) greatly influence the success of imaging experiments in the scanning probe microscopy field. The major goal of the chapter is to provide detailed procedures for sample preparation and operation of the Asylum Research MFP-3D BIO AFM. In addition, one should be cognizant that our comprehensive description in the use of the MFP-3D BIO's functions for successful image acquisitions and analyses is greatly enhanced by Asylum Research's (AR's) accompanying extensive manual(s), technical notes, and AR's users forum. Ultimately, the stepwise protocol and information will allow novice personnel to begin acquiring quality images for processing and analysis with minimal supervision.
authors.affiliation: Department of Pathology and Microscopy Imaging Center, College of Medicine, University of Vermont, Burlington, VT, USA. asquinn@zoo.uvm.edu
authors.forename: Anthony S
authors.initials: AS
authors.lastname: Quinn
authors.forename: Jacob H
authors.initials: JH
authors.lastname: Rand
authors.forename: Xiao-Xuan
authors.initials: XX
authors.lastname: Wu
authors.forename: Douglas J
authors.initials: DJ
authors.lastname: Taatjes
mesh_terms: D000538:Aluminum Silicates / Q000737:chemistry; D017304:Annexin A5 / Q000737:chemistry* / Q000648:ultrastructure; D002021:Buffers; D004791:Enzyme Inhibitors / Q000737:chemistry; D006801:Humans; D006886:Hydroxychloroquine / Q000737:chemistry; D008051:Lipid Bilayers / Q000737:chemistry*; D018625:Microscopy, Atomic Force / Q000295:instrumentation / Q000379:methods*; D010713:Phosphatidylcholines / Q000737:chemistry; D010718:Phosphatidylserines / Q000737:chemistry; D011485:Protein Binding; D053482:beta 2-Glycoprotein I / Q000737:chemistry* / Q000648:ultrastructure
chemical_list: D000538:Aluminum Silicates; D017304:Annexin A5; D002021:Buffers; D004791:Enzyme Inhibitors; D008051:Lipid Bilayers; D010713:Phosphatidylcholines; D010718:Phosphatidylserines; D053482:beta 2-Glycoprotein I; D006886:Hydroxychloroquine; C011934:mica