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.

Atomic force microscopy: High resolution dynamic imaging of cellular and molecular structure in health and disease

“…). To this AnxA5 2D crystal lattice, antibodies from patients with APL, together with the protein cofactor β 2 GPI, were added while simultaneously and temporally imaging with AFM (Quinn et al, ). The resulting images (Fig.…”

“…Accordingly, in the examples from our lab to follow, AC imaging of samples in fluid (biological buffers) was our method of choice to approach molecular imaging of important cell biological/pathobiological questions. The AFM imaging assembly may be used as a stand‐alone instrument, or may be affixed onto a specially designed rigid stage, which in turn is mounted on the base of an inverted light microscope (Quinn et al, ). The immediate advantage of the light microscope component of the system for biomedical samples lies in the ability to optically image the sample and thereby choose microscopically areas of interest for further AFM examination.…”