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Three-dimensional rendering of a large area image of the standard sample. Five identical convoluted images of the tip profile can be seen on the position of each Si spike. The scale bar indicates the dimension in x – y directions. 

Three-dimensional rendering of a large area image of the standard sample. Five identical convoluted images of the tip profile can be seen on the position of each Si spike. The scale bar indicates the dimension in x – y directions. 

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In this work, effective, yet simple, recycling mechanisms for used scanning probe microscopy (SPM) tips were implemented. Comprising a tip profile characterization methodology and specific cleaning procedures, which decontaminate SPM tips whether the contamination nature is known or not, such routes were optimized during numerous tests with brand n...

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... all cleaning agents. The results obtained in this second phase allowed a fine tuning of results obtained in the first phase, ultimately enabling the proposal of a simple recycling route for used SPM tips. Table 1 summarizes the final results after all tests over a hundred ! . Only the most efficient cleaning processes are shown, with their respective optimized cleaning agent combinations and duration times. When inorganic material only is contaminating the SPM tip, the present studies have shown that bathing for 10 s in hydrochloric acid solution 1% v / v, or in hydrofluoric acid solution 1% v / v, followed by 30 s sonication in deionized water are the two most efficient cleaning methods. It must be noted that the HF solution is very aggressive to the SPM tip and eventually damages it if long bathing times are employed. On the other hand, it is very efficient in removing large amounts of inorganic material, and, therefore, is considered an excellent cleaning option when only inorganic contamination is present. Figure 1 illustrates a typical application of HF cleaning. In Figure 1a, a convoluted image of the standard sample indicates the profile of a new SPM tip. The scale bar on the right indicates the SPM tip dimensions in x , y , and z directions. The image in Figure 1b shows the physical profile of the same tip after inorganic material ~ calcium carbonate ! has been purposely aggregated. The enlargement of tip lateral dimensions is evident in this image. Such a tip would normally be considered inadequate for further scans and would usually be discarded. However, after the HF treatment described in Table 1, the tip came out completely clean, as shown in the image of Figure 1c. Actually, it is interesting to note that, in a comparison with Figure 1a, the tip appears to be even sharper than it initially was. It is important to stress that the images in Figure 1a, b, and c are not artifacts due to nonhomogeneities in the standard sample, that is, apart from convolution effects, they represent the true morphology of the SPM tip under investigation. Any effects of nonhomogeneities in the standard sample, like a broken Si spike, are prevented by initially imaging a large area of the sample encompassing several Si spikes and thus producing several convoluted images of the SPM tip. Figure 2 illustrates such a procedure showing, at a different view angle, five images of the tip shown in Figure 1b. Since all images look identical, it is assumed that the standard sample is correctly imaging this SPM tip and, thus, an image of a single Si spike was acquired ~ Fig. 1b ! and used to characterize the tip physical profile. The cleaning efficiency of the HF solution for different types of inorganic contaminants may not be associated with a direct attack on the aggregated material. Rather it seems to be associated with the removal of the silicon oxide layer that always covers a silicon-made SPM tip, either new or used. It is well known in the semiconductor community that HF readily dissolves silicon oxide ~ Ghandhi, 1994 ! . Removing such an oxide layer, where the inorganic contaminants are attached, leads to cleaning of the SPM tip. This oxide removal may also explain the observation that the cleaned tip in Figure 1c appears to be actually sharper than the new tip in Figure 1a. However, such a general and, ...

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... These contaminations may origin from the sample surface or dust in the air. This phenomenon is common in the field of scanning probe microscope (SPM) under ambient conditions, 27 and we usually abandon this probe. To surmount this problem, taking full advantage of the particular structure of our diamond probe, we can separate the diamond chip from the tuning fork handily and clean again with tri-acid or piranha solution (one part of 30% hydrogen peroxide and three parts of 98% concentrated sulfuric acid). ...
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The key component of the scanning magnetometry based on nitrogen-vacancy centers is the diamond probe. Here, we designed and fabricated a new type of probe with an array of pillars on a (100 µm)² × 50 µm diamond chip. The probe features high yield, convertibility to be a single pillar, and expedient reusability. Our fabrication is dramatically simplified by using ultraviolet laser cutting to shape the chip from a diamond substrate instead of additional lithography and time-consuming reactive ion etching. As an example, we demonstrate the imaging of a single magnetic skyrmion with nanoscale resolution. In the future, this flexible probe will be particularly well-suited for commercial applications.
... Thus, imaging setpoints were generally set as low as possible and appropriate values were determined empirically at the beginning of each experiment. The probes were cleaned after each experiment by rinsing in deionized water and acetone (23), and subsequent plasma treatment (PDC 32G; Harrick Plasma, Ithaca, NY) for 10 min. ...
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Contact-mode atomic force microscopy (AFM) has been shown to reveal cortical actin structures. Using live endothelial cells, we visualized cortical actin dynamics simultaneously by AFM and confocal fluorescence microscopy. We present a method that quantifies dynamic changes in the mechanical ultrastructure of the cortical actin web. We argue that the commonly used, so-called error signal imaging in AFM allows a qualitative, but not quantitative, analysis of cortical actin dynamics. The approach we used comprises fast force-curve-based topography imaging and subsequent image processing that enhances local height differences. Dynamic changes in the organization of the cytoskeleton network can be observed and quantified by surface roughness calculations and automated morphometrics. Upon treatment with low concentrations of the actin-destabilizing agent cytochalasin D, the cortical cytoskeleton network is thinned out and the average mesh size increases. In contrast, jasplakinolide, a drug that enhances actin polymerization, consolidates the cytoskeleton network and reduces the average mesh area. In conclusion, cortical actin dynamics can be quantified in live cells. To our knowledge, this opens a new pathway for conducting quantitative structure-function analyses of the endothelial actin web just beneath the apical plasma membrane. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.
Chapter
Tissue elasticity is a critical regulator of cell behavior in normal and diseased conditions like fibrosis and cancer. Since the extracellular matrix (ECM) is a major regulator of tissue elasticity and function, several ECM-based models have emerged in the last decades, including in vitro endogenous ECM, decellularized tissue ECM and ECM hydrogels. The development of such models has urged the need to quantify their elastic properties particularly at the nanometer scale, which is the relevant length scale for cell-ECM interactions. For this purpose, the versatility of atomic force microscopy (AFM) to quantify the nanomechanical properties of soft biomaterials like ECM models has emerged as a very suitable technique. In this chapter we provide a detailed protocol on how to assess the Young's elastic modulus of ECM models by AFM, discuss some of the critical issues, and provide troubleshooting guidelines as well as illustrative examples of AFM measurements, particularly in the context of cancer.