期刊
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
卷 -, 期 118, 页码 -出版社
JOURNAL OF VISUALIZED EXPERIMENTS
DOI: 10.3791/54924
关键词
Engineering; Issue 118; atomic force microscopy; liquid; sub-nanometer resolution imaging; amplitude-modulation; lipid bilayer; biomembranes; crystals; cantilever; harmonics; eigenmode; solvation forces
资金
- Engineering and Physical Sciences Research Council [1452230, EP/M023915/1]
- Biotechnology and Biological Science Research Council [BB/M024830/1]
- European Council [FP7 CIG 631186]
- Biotechnology and Biological Sciences Research Council [BB/M024830/1] Funding Source: researchfish
- Engineering and Physical Sciences Research Council [1788302, 1452230, EP/M023915/1, 1570431] Funding Source: researchfish
- BBSRC [BB/M024830/1] Funding Source: UKRI
- EPSRC [EP/M023915/1, 1788302] Funding Source: UKRI
Atomic force microscopy (AFM) has become a well-established technique for nanoscale imaging of samples in air and in liquid. Recent studies have shown that when operated in amplitude-modulation (tapping) mode, atomic or molecular-level resolution images can be achieved over a wide range of soft and hard samples in liquid. In these situations, small oscillation amplitudes (SAM-AFM) enhance the resolution by exploiting the solvated liquid at the surface of the sample. Although the technique has been successfully applied across fields as diverse as materials science, biology and biophysics and surface chemistry, obtaining high-resolution images in liquid can still remain challenging for novice users. This is partly due to the large number of variables to control and optimize such as the choice of cantilever, the sample preparation, and the correct manipulation of the imaging parameters. Here, we present a protocol for achieving high-resolution images of hard and soft samples in fluid using SAM-AFM on a commercial instrument. Our goal is to provide a step-by-step practical guide to achieving high-resolution images, including the cleaning and preparation of the apparatus and the sample, the choice of cantilever and optimization of the imaging parameters. For each step, we explain the scientific rationale behind our choices to facilitate the adaptation of the methodology to every user's specific system.
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