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.
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Acknowledgement(s)
We are grateful for all the guidance and assistance provided by Asylum Research (http://www.AsylumResearch.com) in our transition to their MFP-3D BIO AFM system. Special thanks to Biological/Applications Scientists Sophia Hohlbach and Nick Geisse; Software Engineers Bruce Wallace, Jason Bemis and Clint Callahan; and Terry Mehr (Director of Marketing Communications), Monte Heaton (EVP Marketing and Business Development) and Genette Dorney (Production Coordinator/Cantilever Sales). This project was supported by Award Number S10RR025498 (to DJT) from the National Center for Research Resources for purchase of the Asylum Research atomic force microscope.
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Quinn, A.S., Rand, J.H., Wu, XX., Taatjes, D.J. (2012). Viewing Dynamic Interactions of Proteins and a Model Lipid Membrane with Atomic Force Microscopy. In: Taatjes, D., Roth, J. (eds) Cell Imaging Techniques. Methods in Molecular Biology, vol 931. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-056-4_13
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DOI: https://doi.org/10.1007/978-1-62703-056-4_13
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