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. 2007 Nov;78(11):113707.
doi: 10.1063/1.2794732.

Quasi-simultaneous imaging/pulling analysis of single polyprotein molecules by atomic force microscopy

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Quasi-simultaneous imaging/pulling analysis of single polyprotein molecules by atomic force microscopy

Alejandro Valbuena et al. Rev Sci Instrum. 2007 Nov.

Abstract

Most of studies in protein nanomechanics have used the atomic force microscope (AFM) in its force-measuring mode on immobilized protein repeats (polyproteins) as single-molecule markers. Here, we add imaging capabilities to a standard, state-of-the-art AFM "puller" and integrate the most powerful programs of analysis available for both AFM modes. This unique instrument allows high-resolution, quasi-simultaneous imaging/force spectroscopy in aqueous solution. We demonstrate its capabilities using polyproteins of a model system (titin I27 domain). This tool should greatly facilitate the development of a much needed universal functionalization system for AFM, one that should allow better sample control and an improved efficiency of protein immobilization.

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