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Summary
Abstract
Introduction
Protocol
Results
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Disclosures
Acknowledgements
Materials
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공학

原子力显微镜悬臂式纳米压痕：空气和流体中纳米级机械性能测量

Published: December 02, 2022
doi:
10.3791/64497
, , , , , , , , , , ,
1Micron School of Materials Science & Engineering,Boise State University, 2Department of Mechanical & Biomedical Engineering,Boise State University, 3Department of Physics,Boise State University, 4Energy and Environmental Science and Technology,Idaho National Laboratory, 5Science and Technology,Idaho National Laboratory, 6Harold Hamm School of Geology & Geological Engineering,University of North Dakota, 7Biomolecular Sciences Graduate Program,Boise State University, 8Center for Advanced Energy Studies

Summary

量化原子力显微镜（AFM）探针尖端施加到样品表面的接触面积和力，可以确定纳米级机械性能。讨论了在软硬样品的空气或流体中实施基于AFM悬臂的纳米压痕的最佳实践，以测量弹性模量或其他纳米力学性能。

Abstract

原子力显微镜（AFM）从根本上测量纳米级AFM探针尖端与样品表面之间的相互作用。如果探针尖端施加的力及其与样品的接触面积可以量化，则可以确定被探测表面的纳米级机械性能（例如，弹性或杨氏模量）。这里提供了进行定量AFM悬臂式纳米压痕实验的详细程序，并提供了如何应用该技术来确定从kPa到GPa的各种样品类型的弹性模量的代表性示例。这些包括生理缓冲液中的活间充质干细胞（MSCs）和细胞核，树脂嵌入的脱水松横截面以及不同成分的Bakken页岩。

此外，基于AFM悬臂的纳米压痕用于探测磷脂双层的断裂强度（即突破力）。讨论了重要的实际考虑因素，例如方法选择和开发、探头选择和校准、感兴趣区域识别、样品异质性、特征尺寸和纵横比、尖端磨损、表面粗糙度以及数据分析和测量统计，以帮助正确实施该技术。最后，证明了AFM衍生的纳米力学图与电子显微镜技术的共定位，该技术提供了有关元素组成的附加信息。

Introduction

了解材料的机械性能是工程中最基本和最重要的任务之一。对于散装材料性能的分析，有许多方法可用于表征材料系统的机械性能，包括拉伸试验1，压缩试验2和三点或四点弯曲（弯曲）试验3。虽然这些微观测试可以提供有关散装材料特性的宝贵信息，但它们通常是在失败的情况下进行的，因此具有破坏性。此外，它们缺乏准确研究当今感兴趣的许多材料系统的微米和纳米级特性所需的空间分辨率，例如薄膜，生物材料和纳米复合材料。为了开始解决大规模机械测试的一些问题，主要是其破坏性，从矿物学中采用了显微硬度测试。硬度是衡量材料在特定条件下对塑性变形的抵抗力的指标。通常，显微硬度测试使用通常由硬化钢或金刚石制成的硬探头压入材料中。然后可以使用所得压痕深度和/或面积来确定硬度。已经开发了几种方法，包括维氏硬度4、努氏硬度 5 和布氏硬度 6;每个都提供了微观材料硬度的测量，但在不同的条件和定义下，因此仅产生可以与在相同条件下进行的测试进行比较的数据。

开发了仪器化纳米压痕，以改进通过各种显微硬度测试方法获得的相对值，提高用于分析机械性能的空间分辨率，并能够分析薄膜。重要的是，通过利用Oliver和Pharr7首先开发的方法，可以通过仪器化纳米压痕确定样品材料的弹性或杨氏模量E。此外，通过使用Berkovich三面锥体纳米压痕探头（其理想的尖端面积函数与维氏四面锥体探头相匹配）8，可以直接比较纳米级和更传统的微米级硬度测量。随着AFM的普及，基于AFM悬臂的纳米压痕也开始受到关注，特别是在测量较软材料的机械性能方面。因此，如图1所示，目前用于询问和量化纳米级机械性能的两种最常用的技术是仪器化纳米压痕（图1A）和基于AFM悬臂的纳米压痕（图1B）9，后者是本工作的重点。

Figure 1
图 1：仪器化和基于 AFM 悬臂的纳米压痕系统的比较。 示意图描述了用于进行（A）仪器化纳米压痕和（B）基于AFM悬臂的纳米压痕的典型系统。这个数字是从钱等人51修改而来的。缩写：AFM = 原子力显微镜。 请点击此处查看此图的大图。

仪器化和基于AFM悬臂的纳米压痕都使用刚性探针使感兴趣的样品表面变形，并监测合力和位移作为时间的函数。通常，所需的负载（即力）或（Z压电）位移曲线由用户 通过 软件界面指定并由仪器直接控制，而另一个参数则被测量。最常从纳米压痕实验中获得的机械性能是弹性模量（E），也称为杨氏模量，它具有压力单位。材料的弹性模量是与粘结刚度有关的基本属性，定义为在塑性变形开始之前，拉伸或压应力（σ，每单位面积施加的力）与轴向应变（ε，沿压痕轴的比例变形）之比（即可逆或暂时）变形（方程 [1]）：

Equation 1（1）

应该注意的是，由于许多材料（特别是生物组织）实际上是粘弹性的，因此实际上，（动态或复杂）模量由弹性（存储，同相）和粘性（损失，异相）成分组成。在实际操作中，在纳米压痕实验中测量的是还原模量 E*，它与目标的真实样品模量 E有关，如公式（2）所示：

Equation 2（二）

其中 E 尖端和 ν 尖端分别是纳米压痕尖端的弹性模量和泊松比，ν 是样品的估计泊松比。泊松比是横向应变与轴向应变的负比，因此表示样品在承受轴向应变时（例如，在纳米压痕载荷期间）的横向伸长程度，如公式（3）所示：

Equation 3（三）

从降低模量转换为实际模量是必要的，因为a）压头尖端施加的一些轴向应变可以转换为横向应变（即，样品可能通过垂直于加载方向的膨胀或收缩而变形），以及b）压头不是无限硬的，因此压痕样品的行为导致尖端的一些（小）变形量。请注意，在E尖端>>E的情况下（即压头比样品硬得多，这在使用金刚石探头时通常是正确的），减少的样品模量和实际样品模量之间的关系大大简化为E ≈ E*（1 – v2）。 虽然仪器化纳米压痕在准确的力表征和动态范围方面具有优势，但基于AFM悬臂的纳米压痕速度更快，提供更高数量级的力和位移灵敏度，可实现更高分辨率的成像和改进的压痕定位，并且可以同时探测纳米级的磁性和电学特性9.特别是，基于AFM悬臂的纳米压痕在纳米级软材料（例如，聚合物，凝胶，脂质双层和细胞或其他生物材料），极薄（亚μm）薄膜（基底效应可能取决于压痕深度）10，11和悬浮二维（2D）材料12，13，14（例如石墨烯）的纳米级机械性能方面具有优越性15，16，云母17，六方氮化硼（h-BN）18或过渡金属硫族化物（TMDC;例如MoS2）19。这是由于其精细的力（sub-nN）和位移（sub-nm）灵敏度，这对于准确确定初始接触点并保持在弹性变形区域内非常重要。

在基于AFM悬臂的纳米压痕中，AFM探针向样品表面的位移由校准的压电元件驱动（图1B），柔性悬臂最终由于与样品表面接触时经历的电阻力而弯曲。悬臂的这种弯曲或偏转通常通过将激光从悬臂背面反射到光电探测器（位置敏感探测器 [PSD]）中来监测。再加上悬臂刚度（以nN / nm为单位）和偏转灵敏度（以nm/V为单位）的知识，可以将测量的悬臂挠度（以V为单位）转换为施加在样品上的力（以nN为单位）。接触后，Z-压电运动和悬臂偏转之间的差异会产生样品压痕深度。结合尖端面积功能的知识，可以计算尖端-样品接触面积。然后可以使用适当的接触力学模型（请参阅讨论的 数据分析 部分）拟合所得力-距离或力-位移（F-D）曲线的接触部分的斜率，以确定样品的纳米力学性能。虽然基于AFM悬臂的纳米压痕与上述仪器化纳米压痕相比具有一些明显的优势，但它也带来了一些实际实施挑战，例如校准，尖端磨损和数据分析，本文将对此进行讨论。基于AFM悬臂的纳米压痕的另一个潜在缺点是假设线性弹性，因为接触半径和压痕深度需要远小于压头半径，这在使用纳米级AFM探头和/或表现出显着表面粗糙度的样品时可能难以实现。

传统上，纳米压痕仅限于单个位置或小网格压痕实验，其中选择所需的位置（即感兴趣区域[ROI]）并且单个受控压痕，单个位置中的多个压痕由一些等待时间隔开，和/或以Hz量级的速率执行缩进的粗网格。然而，AFM的最新进展允许通过使用基于高速力曲线的成像模式（根据系统制造商的不同商品名称）同时获取机械性能和形貌，其中力曲线在负载控制下以kHz速率进行，最大尖端样品力用作成像设定点。还开发了傻瓜式方法，允许获取AFM形貌图像，然后在图像内的兴趣点进行选择性纳米压痕，从而对纳米压痕位置进行纳米级空间控制。虽然不是这项工作的主要重点，但代表性结果中提供了基于力曲线的成像和基于傻瓜悬臂的纳米压痕的特定 选定应用示例 ，并且可以与下面概述的协议结合使用（如果在采用的特定AFM平台上可用）。具体来说，这项工作概述了在任何有能力的AFM系统上实际实施基于AFM悬臂的纳米压痕的通用协议，并提供了该技术的四个用例示例（两个在空气中，两个在流体中），包括代表性结果和对细微差别的深入讨论，挑战和成功使用该技术的重要考虑因素。

Protocol

注意：由于商用AFM种类繁多，并且基于悬臂的纳米压痕存在多种样品类型和应用，因此以下方案有意设计为相对通用的性质，侧重于所有悬臂式纳米压痕实验所需的共享步骤，无论仪器或制造商如何。正因为如此，作者假设读者至少对操作用于执行悬臂式纳米压痕的特定仪器具有基本的熟悉程度。然而，除了下面概述的一般协议外，还包括特定于AFM和此处使用的软件的详细分步标准操作程序（SOP?…

Representative Results

力-位移曲线图 7 显示了在树脂嵌入的枇杷松样品（图 7A）和间充质干细胞 （MSC） 细胞核上的液体（磷酸盐缓冲盐水 [PBS]）上进行的纳米压痕实验中获得的具有代表性的、近乎理想的 F-D 曲线（图 7B）。任何接触力学模型的使用都依赖于对初始针尖-样品接触点的准确可靠测定。因此，初始接触点之前相对平坦、低?…

Discussion

样品制备
对于空气中的纳米压痕，常见的制备方法包括冷冻切片（例如，组织样品）、研磨和/或抛光，然后进行超薄切片（例如，树脂嵌入的生物样品）、离子研磨或聚焦离子束制备（例如，不适合抛光的半导体、多孔或混合硬度样品）、机械或电化学抛光（例如，金属合金）或薄膜沉积（例如，原子层或化学气相沉积， 分子束外延）。目标是创建具有最小表面粗糙度的样品（?…

Disclosures

The authors have nothing to disclose.

Acknowledgements

所有AFM实验均在博伊西州立大学表面科学实验室（SSL）进行。SEM表征在博伊西州立材料表征中心（BSCMC）进行。本出版物中报道的有关生物燃料原料的研究得到了美国能源部、能源效率和可再生能源办公室、生物能源技术办公室作为原料转换接口联盟 （FCIC） 的一部分以及美国能源部爱达荷州运营办公室合同 DE-AC07-051ID14517 的部分支持。细胞力学研究得到了美国国立卫生研究院（美国）AG059923、AR075803和P20GM109095的资助，以及美国国家科学基金会（美国）的1929188和2025505资助。模型脂质双层系统工作得到了美国国立卫生研究院（美国）R01 EY030067资助的支持。作者感谢Elton Graugnard博士制作了 图11所示的合成图像。

Materials

Atomic force microscope Bruker Dimension Icon Uses Nanoscope control software, including PeakForce Quantitative Nanomechanical Mapping (PF-QNM), FastForce Volume (FFV), and Point-and-Shoot Ramping experimental workspaces
AtomicJ American Institute of Physics https://doi.org/10.1063/1.4881683 Flexible, powerful, free open source Java-based force curve analysis software package. Supports numerous contact mechanic models, such as Hertz, Sneddon DMT, JKR, Maugis, and cone or pyramid (including blunt and truncated). Also includes a variety of initial contact point estimation methods to choose from. Supports batch processing of data and subsequent statistical analysis (e.g., averages, standard deviations, histograms, goodness of fit, etc.). Literature citation is: P. Hermanowicz, M. Sarna, K. Burda, and H. GabryEquation 1, “AtomicJ: An open source software for analysis of force curves” Rev. Sci. Instrum. 85: 063703 (2014), https://doi.org/10.1063/1.4881683
Buffer solution (PBS) Fisher Chemical (NaCl), Sigma Aldrich (KCl), Fisher BioReagents (Na2HPO4 and KH2PO4) S271 (>99% purity NaCl), P9541 (>99% purity KCl), BP332(>99% purity Na2HPO4), BP362 (>99% purity KH2PO4) Phosphate buffered saline (PBS) was prepared in the laboratory as an aqueous solution consisting of 137 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, and 1.8 mM KH2PO4 dissolved in ultrapure water. Reagents were measured out using an analytical balance, and glassware was cleaned with soap and water followed by autoclaving immediately prior to use.
Chloroform
Diamond tip AFM probe Bruker PDNISP Pre-mounted factory-calibrated cube corner diamond (E = 1140 GPa) tip AFM probe (nominal R = 40 nm) with a stainless steel cantilever (nominal k = 225 N/m, f0 = 50 kHz). Spring constant is measured at the factory (k = 256 N/m for the probe, Serial #13435414, used here) and calibration data (including AFM images of indents showing probe geometry) is provided with the probe.
Diamond ultramicrotome blade Diatome Ultra 35° 2.1 mm width. Also used a standard glass blade for intial rough cut of sample surface before transitioning to diamond blade for final surface preparation
Epoxy Gorilla Glue 26853-31-6 Epoxy resin and hardner were mixed in a 1:1 ratio, a small drop was placed on a stainless steel sample puck (Ted Pella), and V1 grade muscovite mica (Ted Pella) was attached to create an atomically flat surface for preparation of phospholipid membranes.
Ethanol
LR white resin, medium grade (catalyzed) Electron Microscopy Sciences 14381 500 mL bottle, Lot #150629
Mesenchymal stem cells (MSCs) N/A N/A MSCs for nanomechanical studies were primary cells harvested from 8-10 week old male C57BL/6 mice as described in Goelzer, M. et al. "Lamin A/C Is Dispensable to Mechanical Repression of Adipogenesis" Int J Mol Sci 22: 6580 (2021) doi:10.3390/ijms22126580 and Peister, A. et al. "Adult stem cells from bone marrow (MSCs) isolated from different strains of inbred mice vary in surface epitopes, rates of proliferation, and differentiation potential" Blood 103: 1662-1668 (2004), doi:10.1182/blood-2003-09-3070.
Modulus standards Bruker PFQNM-SMPKIT-12M Used HOPG (E = 18 GPa) and PS (E = 2.7 GPa). Also contains 2x PDMS (Tack 0, E = 2.5 MPa; Tack 4, E = 3.5 MPa), PS-LDPE (E = 2.0/0.2 GPa), fused silica (E = 72.9 GPa), sapphire (E – 345 GPa), and tip characterization (titanium roughness) sample. All samples come pre-mounted on a 12 mm diameter steel disc (sample puck).
Muscovite mica Ted Pella 50-12 12 mm diameter, V1 grade muscovite mica
Nanscope Analysis Bruker Version 2.0 Free AFM image processing and analysis software package, but designed for, and proprietary/limited to Bruker AFMs; similar functionality is available from free, platform-independent AFM image processing and analysis software packages such as Gwyddion, WSxM, and others. Has built-in capabilities for force curve analysis, but AtomicJ is more flexible/full featured (e.g., more built-in contact mechanics models to choose from, statistical analysis of force curve fitting results, etc.) for force curve analysis and handles batch processing of force curves.
Phospholipids: POPC, Cholesterol (ovine) Avanti Polar Lipids POPC: CAS # 26853-31-6, Cholesterol: CAS # 57-88-5 POPC lipid dissolved in chloroform (25 mg/mL) was obtained from vendor and used without further purification. Cholesterol powder from the same vendor was dissolved in chloroform (20 mg/mL). 
Probe holder (fluid, lipid bilayers) Bruker MTFML-V2 Specific to the particular AFM used; MTFML-V2 is a glass probe holder for scanning in fluid on a MultiMode AFM.
Probe holder (fluid, MSCs) Bruker FastScan Bio Z-scanner Used with Dimension FastScan head (XY flexure scanners). Serial number MXYPOM5-1B154.
Probe holder (standard, ambient) Bruker DAFMCH Specific to the particular AFM used; DAFMCH is the standard contact and tapping mode probe holder for the Dimension Icon AFM, suitable for nanoindentation (PF-QNM, FFV, and point-and-shoot ramping)
Sample Puck Ted Pella 16218 Product number is for 15 mm diameter stainless steel sample puck. Also available in 6 mm, 10 mm, 12 mm, and 20 mm diameters at https://www.tedpella.com/AFM_html/AFM.aspx#anchor842459
Sapphire substrate Bruker PFQNM-SMPKIT-12M Extremely hard surface (E = 345 GPa) for measuring deflection sensitivity of probes (want all of the deflection to come from the probe, not the substrate). Part of the PF-QNM/modulus standards kit.
Scanning electron microscope Hitachi S-3400N-II Located at Boise State. Used to perform co-localized SEM/EDS on all samples except additively manufactured (AM) Ti-6Al-4V.
Silicon AFM probes (standard) NuNano Scout 350 Standard tapping mode silicon probe with reflective aluminum backside coating; k = 42 N/m (nominal), f0 = 350 kHz. Nominal R = 5 nm. Also available uncoated or with reflective gold backside coating. Probes with similar specifications are available from other manufacturers (e.g., Bruker TESPA-V2).
Silicon AFM probes (stiff) Bruker RTESPA-525, RTESPA-525-30  Rotated tip etched silicon probes with reflective aluminum backside coating; k = 200 N/m (nominal), f0 = 525 kHz. Nominal R = 8 nm for RTESPA-525, R = 30 nm for RTESPA-525-30. Spring constant of each RTESPA-525-30 is measured individually at the factory via laser Doppler vibrometry and supplied with the probe.
Silicon carbide grit paper (abrasive discs) Allied 50-10005 120 grit
Silicon nitride AFM probes (soft, large radius hemispherical tip) Bruker MLCT-SPH-5UM, MLCT-SPH-5UM-DC Also MLCT-SPH-1UM-DC. New product line of factory-calibrated (probe radius and spring constants of all cantilevers) large radius (R = 1 or 5 mm) hemispherical tip (at the end of a 23 mm long cylindrical shaft) probes. DC = drift compensation coating. 6 cantilevers/probe (A-F). Nominal spring constants: A, k = 0.07 N/m; B, k = 0.02 N/m; C, k = 0.01 N/m; D, k = 0.03 N/m; E, k = 0.1 N/m; F, k = 0.6 N/m.
Silicon nitride AFM probes (soft, medium sharp tip) Bruker DNP 4 cantilevers/probe (A-d). Nominal spring constants: A, k = 0.35 N/m; B, k = 0.12 N/m; C, k = 0.24 N/m; D, k = 0.06 N/m. Nominal radii of curvature, R = 10 nm.
Silicon nitride AFM probes (soft, sharp tip) Bruker ScanAsyst-Air Nominal values: resonance frequency, f0 = 70 kHz; spring constant, k = 0.4 N/m; radius of curvature, R = 2 nm. Designed for force curve based AFM imaging.
Superglue Henkel Loctite 495 Cyanoacrylate based instant adhesive. Lots of roughly equivalent products are readily available.
Syringe pump New Era Pump Systems NE1000US One channel syringe pump system with infusion and withdrawal capacity
Tip characterization standard Bruker PFQNM-SMPKIT-12M Titanium (Ti) roughness standard. Part of the PF-QNM/modulus standards kit.
Ultrahigh purity nitrogen (UHP N2), 99.999% Norco SPG TUHPNI – T T size compressed gas cylinder of ultrahigh purity (99.999%) nitrogen for drying samples
Ultramicrotome Leica EM UC6 Equipped with a glass blade (standard, for intial sample preparation) and a diamond blade (for final preparation)
Ultrapure water Thermo Fisher Barnstead Nanopure Model 7146 Model has been discontinued, but equivalent products are available. Produces ≥18.2 MΩ*cm ultrapure water with 1-5 ppb TOC (total organic content), per inline UV monitoring. Includes 0.2 µm particulate filter, ion exchange columns, and UV oxidation chamber.
Variable Speed Grinder Buehler EcoMet 3000 Used with silicon carbide grit papers during hand polishing.
Vibration isolation table (active) Herzan TS-140 Used with Bruker MultiMode AFM. Sits on a TMC 65-531 vibration isolation table. Bruker Dimension Icon AFM utilizes strictly passive vibration isolation (comes from manufacturer with custom acoustic hood, air table, and granite slab).
Vibration isolation table (passive) TMC 65-531 35" x 30" vibration isolation table with optional air damping (disabled). Used with Bruker MultiMode AFM. Herzan TS-140 "Table Stable" active vibration control table is located on top.
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References

  1. Hart, E. W. Theory of the tensile test. Acta Metallurgica. 15 (2), 351-355 (1967).
  2. Fell, J. T., Newton, J. M. Determination of tablet strength by the diametral-compression test. Journal of Pharmaceutical Sciences. 59 (5), 688-691 (1970).
  3. Babiak, M., Gaff, M., Sikora, A., Hysek, &. #. 3. 5. 2. ;. Modulus of elasticity in three- and four-point bending of wood. Composite Structures. 204, 454-465 (2018).
  4. Song, S., Yovanovich, M. M. Relative contact pressure-Dependence on surface roughness and Vickers microhardness. Journal of Thermophysics and Heat Transfer. 2 (1), 43-47 (1988).
  5. Hays, C., Kendall, E. G. An analysis of Knoop microhardness. Metallography. 6 (4), 275-282 (1973).
  6. Hill, R., Storåkers, B., Zdunek, A. B. A theoretical study of the Brinell hardness test. Proceedings of the Royal Society of London. A. Mathematical and Physical Sciences. 423 (1865), 301-330 (1989).
  7. Oliver, W. C., Pharr, G. M. An improved technique for determining hardness and elastic modulus using load and displacement sensing indentation experiments. Journal of Materials Research. 7 (6), 1564-1583 (1992).
  8. Sakharova, N. A., Fernandes, J. V., Antunes, J. M., Oliveira, M. C. Comparison between Berkovich, Vickers and conical indentation tests: A three-dimensional numerical simulation study. International Journal of Solids and Structures. 46 (5), 1095-1104 (2009).
  9. Cohen, S. R., Kalfon-Cohen, E. Dynamic nanoindentation by instrumented nanoindentation and force microscopy: a comparative review. Beilstein Journal of Nanotechnology. 4 (1), 815-833 (2013).
  10. Saha, R., Nix, W. D. Effects of the substrate on the determination of thin film mechanical properties by nanoindentation. Acta Materialia. 50 (1), 23-38 (2002).
  11. Tsui, T. Y., Pharr, G. M. Substrate effects on nanoindentation mechanical property measurement of soft films on hard substrates. Journal of Materials Research. 14 (1), 292-301 (1999).
  12. Cao, G., Gao, H. Mechanical properties characterization of two-dimensional materials via nanoindentation experiments. Progress in Materials Science. 103, 558-595 (2019).
  13. Castellanos-Gomez, A., Singh, V., vander Zant, H. S. J., Steele, G. A. Mechanics of freely-suspended ultrathin layered materials. Annalen der Physik. 527 (1-2), 27-44 (2015).
  14. Cao, C., Sun, Y., Filleter, T. Characterizing mechanical behavior of atomically thin films: A review. Journal of Materials Research. 29 (3), 338-347 (2014).
  15. Lee, C., Wei, X., Kysar, J. W., Hone, J. Measurement of the elastic properties and intrinsic strength of monolayer graphene. Science. 321 (5887), 385-388 (2008).
  16. Elibol, K., et al. Visualising the strain distribution in suspended two-dimensional materials under local deformation. Scientific Reports. 6 (1), 28485 (2016).
  17. Castellanos-Gomez, A., et al. Mechanical properties of freely suspended atomically thin dielectric layers of mica. Nano Research. 5 (8), 550-557 (2012).
  18. Song, L., et al. Large scale growth and characterization of atomic hexagonal boron nitride layers. Nano Letters. 10 (8), 3209-3215 (2010).
  19. Castellanos-Gomez, A., et al. Elastic properties of freely suspended MoS2 nanosheets. Advanced Materials. 24 (6), 772-775 (2012).
  20. D’Costa, N. P., Hoh, J. H. Calibration of optical lever sensitivity for atomic force microscopy. Review of Scientific Instruments. 66 (10), 5096-5097 (1995).
  21. Wu, Y., et al. Evaluation of elastic modulus and hardness of crop stalks cell walls by nano-indentation. Bioresource Technology. 101 (8), 2867-2871 (2010).
  22. Barns, S., et al. Investigation of red blood cell mechanical properties using AFM indentation and coarse-grained particle method. BioMedical Engineering OnLine. 16 (1), 140 (2017).
  23. Hermanowicz, P., Sarna, M., Burda, K., Gabryś, H. AtomicJ: An open source software for analysis of force curves. Review of Scientific Instruments. 85 (6), 063703 (2014).
  24. Broitman, E. Indentation hardness measurements at macro-, micro-, and nanoscale: a critical overview. Tribology Letters. 65 (1), 23 (2016).
  25. Tiwari, A. . Nanomechanical Analysis of High Performance Materials. , (2015).
  26. Aggarwal, R. L., Ramdas, A. K. . Physical Properties of Diamond and Sapphire. , (2019).
  27. Boyd, E. J., Uttamchandani, D. Measurement of the anisotropy of Young’s modulus in single-crystal silicon. Journal of Microelectromechanical Systems. 21 (1), 243-249 (2012).
  28. Harding, J. W., Sneddon, I. N. The elastic stresses produced by the indentation of the plane surface of a semi-infinite elastic solid by a rigid punch. Mathematical Proceedings of the Cambridge Philosophical Society. 41 (1), 16-26 (2008).
  29. Lin, D. C., Dimitriadis, E. K., Horkay, F. Robust strategies for automated AFM force curve analysis-I. Non-adhesive indentation of soft, inhomogeneous materials. Journal of Biomechanical Engineering. 129 (3), 430-440 (2006).
  30. Lin, D. C., Dimitriadis, E. K., Horkay, F. Robust strategies for automated AFM force curve analysis-II: Adhesion-influenced indentation of soft, elastic materials. Journal of Biomechanical Engineering. 129 (6), 904-912 (2007).
  31. Haile, S., Palmer, M., Otey, A. Potential of loblolly pine: switchgrass alley cropping for provision of biofuel feedstock. Agroforestry Systems. 90 (5), 763-771 (2016).
  32. Lu, X., et al. Biomass logistics analysis for large scale biofuel production: Case study of loblolly pine and switchgrass. Bioresource Technology. 183, 1-9 (2015).
  33. Susaeta, A., Lal, P., Alavalapati, J., Mercer, E., Carter, D. Economics of intercropping loblolly pine and switchgrass for bioenergy markets in the southeastern United States. Agroforestry Systems. 86 (2), 287-298 (2012).
  34. Garcia, R. Nanomechanical mapping of soft materials with the atomic force microscope: methods, theory and applications. Chemical Society Reviews. 49 (16), 5850-5884 (2020).
  35. Derjaguin, B. V., Muller, V. M., Toporov, Y. P. Effect of contact deformations on the adhesion of particles. Journal of Colloid and Interface Science. 53 (2), 314-326 (1975).
  36. Ciesielski, P. N., et al. Engineering plant cell walls: tuning lignin monomer composition for deconstructable biofuel feedstocks or resilient biomaterials. Green Chemistry. 16 (5), 2627-2635 (2014).
  37. Liu, K., Ostadhassan, M., Zhou, J., Gentzis, T., Rezaee, R. Nanoscale pore structure characterization of the Bakken shale in the USA. Fuel. 209, 567-578 (2017).
  38. Maryon, O. O., et al. Co-localizing Kelvin probe force microscopy with other microscopies and spectroscopies: selected applications in corrosion characterization of alloys. JoVE. (184), e64102 (2022).
  39. Eliyahu, M., Emmanuel, S., Day-Stirrat, R. J., Macaulay, C. I. Mechanical properties of organic matter in shales mapped at the nanometer scale. Marine and Petroleum Geology. 59, 294-304 (2015).
  40. Li, C., et al. Nanomechanical characterization of organic matter in the Bakken formation by microscopy-based method. Marine and Petroleum Geology. 96, 128-138 (2018).
  41. Bouzid, T., et al. The LINC complex, mechanotransduction, and mesenchymal stem cell function and fate. Journal of Biological Engineering. 13 (1), 68 (2019).
  42. Dupont, S., et al. Role of YAP/TAZ in mechanotransduction. Nature. 474 (7350), 179-183 (2011).
  43. Wang, S., et al. CCM3 is a gatekeeper in focal adhesions regulating mechanotransduction and YAP/TAZ signalling. Nature Cell Biology. 23 (7), 758-770 (2021).
  44. Sen, B., et al. Mechanical strain inhibits adipogenesis in mesenchymal stem cells by stimulating a durable β-catenin signal. Endocrinology. 149 (12), 6065-6075 (2008).
  45. Sen, B., et al. mTORC2 regulates mechanically induced cytoskeletal reorganization and lineage selection in marrow-derived mesenchymal stem cells. Journal of Bone and Mineral Research. 29 (1), 78-89 (2014).
  46. Sen, B., et al. Mechanically induced nuclear shuttling of β-catenin requires co-transfer of actin. Stem Cells. 40 (4), 423-434 (2022).
  47. Newberg, J., et al. Isolated nuclei stiffen in response to low intensity vibration. Journal of Biomechanics. 111, 110012 (2020).
  48. Ding, Y., Xu, G. -. K., Wang, G. -. F. On the determination of elastic moduli of cells by AFM based indentation. Scientific Reports. 7 (1), 45575 (2017).
  49. Khadka, N. K., Timsina, R., Rowe, E., O’Dell, M., Mainali, L. Mechanical properties of the high cholesterol-containing membrane: An AFM study. Biochimica et Biophysica Acta. Biomembranes. 1863 (8), 183625 (2021).
  50. Castellana, E. T., Cremer, P. S. Solid supported lipid bilayers: From biophysical studies to sensor design. Surface Science Reports. 61 (10), 429-444 (2006).
  51. Qian, L., Zhao, H. Nanoindentation of soft biological materials. Micromachines. 9 (12), 654 (2018).
  52. Pittenger, B., Yablon, D. Improving the accuracy of nanomechanical measurements with force-curve-based AFM techniques. Bruker Application Notes. 149, (2017).
  53. Vorselen, D., Kooreman, E. S., Wuite, G. J. L., Roos, W. H. Controlled tip wear on high roughness surfaces yields gradual broadening and rounding of cantilever tips. Scientific Reports. 6 (1), 36972 (2016).
  54. Bhaskaran, H., et al. Ultralow nanoscale wear through atom-by-atom attrition in silicon-containing diamond-like carbon. Nature Nanotechnology. 5 (3), 181-185 (2010).
  55. Giannazzo, F., Schilirò, E., Greco, G., Roccaforte, F. Conductive atomic force microscopy of semiconducting transition metal dichalcogenides and heterostructures. Nanomaterials. 10 (4), 803 (2020).
  56. Melitz, W., Shen, J., Kummel, A. C., Lee, S. Kelvin probe force microscopy and its application. Surface Science Reports. 66 (1), 1-27 (2011).
  57. Kazakova, O., et al. Frontiers of magnetic force microscopy. Journal of Applied Physics. 125 (6), 060901 (2019).
  58. Kim, H. -. J., Yoo, S. -. S., Kim, D. -. E. Nano-scale wear: A review. International Journal of Precision Engineering and Manufacturing. 13 (9), 1709-1718 (2012).
  59. Heath, G. R., et al. Localization atomic force microscopy. Nature. 594 (7863), 385-390 (2021).
  60. Strahlendorff, T., Dai, G., Bergmann, D., Tutsch, R. Tip wear and tip breakage in high-speed atomic force microscopes. Ultramicroscopy. 201, 28-37 (2019).
  61. Lantz, M. A., et al. Wear-resistant nanoscale silicon carbide tips for scanning probe applications. Advanced Functional Materials. 22 (8), 1639-1645 (2012).
  62. Khurshudov, A. G., Kato, K., Koide, H. Wear of the AFM diamond tip sliding against silicon. Wear. 203, 22-27 (1997).
  63. Villarrubia, J. S. Algorithms for scanned probe microscope image simulation, surface reconstruction, and tip estimation. Journal of Research of the National Institute of Standards and Technology. 102 (4), 425 (1997).
  64. Kain, L., et al. Calibration of colloidal probes with atomic force microscopy for micromechanical assessment. Journal of the Mechanical Behavior of Biomedical Materials. 85, 225-236 (2018).
  65. Slattery, A. D., Blanch, A. J., Quinton, J. S., Gibson, C. T. Accurate measurement of Atomic Force Microscope cantilever deflection excluding tip-surface contact with application to force calibration. Ultramicroscopy. 131, 46-55 (2013).
  66. Dobrovinskaya, E. R., Lytvynov, L. A., Pishchik, V. . Sapphire: Material, Manufacturing, Applications. , (2009).
  67. te Riet, J., et al. Interlaboratory round robin on cantilever calibration for AFM force spectroscopy. Ultramicroscopy. 111 (12), 1659-1669 (2011).
  68. Pratt, J. R., Shaw, G. A., Kumanchik, L., Burnham, N. A. Quantitative assessment of sample stiffness and sliding friction from force curves in atomic force microscopy. Journal of Applied Physics. 107 (4), 044305 (2010).
  69. Slattery, A. D., Blanch, A. J., Quinton, J. S., Gibson, C. T. Calibration of atomic force microscope cantilevers using standard and inverted static methods assisted by FIB-milled spatial markers. Nanotechnology. 24 (1), 015710 (2012).
  70. Higgins, M. J., et al. Noninvasive determination of optical lever sensitivity in atomic force microscopy. Review of Scientific Instruments. 77 (1), 013701 (2006).
  71. Lévy, R., Maaloum, M. Measuring the spring constant of atomic force microscope cantilevers: thermal fluctuations and other methods. Nanotechnology. 13 (1), 33-37 (2001).
  72. Sikora, A. Quantitative normal force measurements by means of atomic force microscopy towards the accurate and easy spring constant determination. Nanoscience and Nanometrology. 2 (1), 8-29 (2016).
  73. Ohler, B. Cantilever spring constant calibration using laser Doppler vibrometry. Review of Scientific Instruments. 78 (6), 063701 (2007).
  74. Gates, R. S., Pratt, J. R. Accurate and precise calibration of AFM cantilever spring constants using laser Doppler vibrometry. Nanotechnology. 23 (37), 375702 (2012).
  75. Cleveland, J. P., Manne, S., Bocek, D., Hansma, P. K. A nondestructive method for determining the spring constant of cantilevers for scanning force microscopy. Review of Scientific Instruments. 64 (2), 403-405 (1993).
  76. Sader, J. E., Chon, J. W. M., Mulvaney, P. Calibration of rectangular atomic force microscope cantilevers. Review of Scientific Instruments. 70 (10), 3967-3969 (1999).
  77. Sader, J. E., et al. Spring constant calibration of atomic force microscope cantilevers of arbitrary shape. Review of Scientific Instruments. 83 (10), 103705 (2012).
  78. Sader, J. E. Frequency response of cantilever beams immersed in viscous fluids with applications to the atomic force microscope. Journal of Applied Physics. 84 (1), 64-76 (1998).
  79. Sader, J. E., Pacifico, J., Green, C. P., Mulvaney, P. General scaling law for stiffness measurement of small bodies with applications to the atomic force microscope. Journal of Applied Physics. 97 (12), 124903 (2005).
  80. Mendels, D. -. A., et al. Dynamic properties of AFM cantilevers and the calibration of their spring constants. Journal of Micromechanics and Microengineering. 16 (8), 1720-1733 (2006).
  81. Gao, S., Brand, U. In-situ nondestructive characterization of the normal spring constant of AFM cantilevers. Measurement Science and Technology. 25 (4), 044014 (2014).
  82. Gibson, C. T., Watson, G. S., Myhra, S. Determination of the spring constants of probes for force microscopy/spectroscopy. Nanotechnology. 7 (3), 259-262 (1996).
  83. Gates, R. S., Pratt, J. R. Prototype cantilevers for SI-traceable nanonewton force calibration. Measurement Science and Technology. 17 (10), 2852-2860 (2006).
  84. Neumeister, J. M., Ducker, W. A. Lateral, normal, and longitudinal spring constants of atomic force microscopy cantilevers. Review of Scientific Instruments. 65 (8), 2527-2531 (1994).
  85. Kim, M. S., Choi, I. M., Park, Y. K., Kang, D. I. Atomic force microscope probe calibration by use of a commercial precision balance. Measurement. 40 (7), 741-745 (2007).
  86. Kim, M. -. S., Choi, J. -. H., Park, Y. -. K., Kim, J. -. H. Atomic force microscope cantilever calibration device for quantified force metrology at micro- or nano-scale regime: the nano force calibrator (NFC). Metrologia. 43 (5), 389-395 (2006).
  87. Tian, Y., et al. A novel method and system for calibrating the spring constant of atomic force microscope cantilever based on electromagnetic actuation. Review of Scientific Instruments. 89 (12), 125119 (2018).
  88. Clifford, C. A., Seah, M. P. The determination of atomic force microscope cantilever spring constants via dimensional methods for nanomechanical analysis. Nanotechnology. 16 (9), 1666-1680 (2005).
  89. Chen, B. -. Y., Yeh, M. -. K., Tai, N. -. H. Accuracy of the spring constant of atomic force microscopy cantilevers by finite element method. Analytical Chemistry. 79 (4), 1333-1338 (2007).
  90. Mick, U., Eichhorn, V., Wortmann, T., Diederichs, C., Fatikow, S. Combined nanorobotic AFM/SEM system as novel toolbox for automated hybrid analysis and manipulation of nanoscale objects. 2010 IEEE International Conference on Robotics and Automation. , 4088-4093 (2010).
  91. Kim, M. -. S., Choi, J. -. H., Kim, J. -. H., Park, Y. -. K. Accurate determination of spring constant of atomic force microscope cantilevers and comparison with other methods. Measurement. 43 (4), 520 (2010).
  92. Zhang, G., Wei, Z., Ferrell, R. E. Elastic modulus and hardness of muscovite and rectorite determined by nanoindentation. Applied Clay Science. 43 (2), 271-281 (2009).
  93. Bobko, C. P., Ortega, J. A., Ulm, F. -. J. Comment on "Elastic modulus and hardness of muscovite and rectorite determined by nanoindentation by G. Zhang, Z. Wei and R.E. Ferrell. Applied Clay Science. 46 (4), 425-428 (2009).
  94. Zhang, G., Wei, Z., Ferrell, R. E. Reply to the Comment on "Elastic modulus and hardness of muscovite and rectorite determined by nanoindentation" by G. Zhang, Z. Wei and R. E. Ferrell. Applied Clay Science. 46 (4), 429-432 (2009).
  95. Jin, D. W., et al. Thermal stability and Young’s modulus of mechanically exfoliated flexible mica. Current Applied Physics. 18 (12), 1486-1491 (2018).
  96. Xiao, J., et al. Anisotropic friction behaviour of highly oriented pyrolytic graphite. Carbon. 65, 53-62 (2013).
  97. Hertz, H. Ueber die Berührung fester elastischer Körper. Journal für die reine und angewandte Mathematik. 1882 (92), 156-171 (1882).
  98. Johnson, K. L., Kendall, K., Roberts, A. D., Tabor, D. Surface energy and the contact of elastic solids. Proceedings of the Royal Society of London. A. Mathematical and Physical Sciences. 324 (1558), 301-313 (1971).
  99. Muller, V. M., Derjaguin, B. V., Toporov, Y. P. On two methods of calculation of the force of sticking of an elastic sphere to a rigid plane. Colloids and Surfaces. 7 (3), 251-259 (1983).
  100. Maugis, D. Adhesion of spheres: The JKR-DMT transition using a dugdale model. Journal of Colloid and Interface Science. 150 (1), 243-269 (1992).
  101. Muller, V. M., Yushchenko, V. S., Derjaguin, B. V. On the influence of molecular forces on the deformation of an elastic sphere and its sticking to a rigid plane. Journal of Colloid and Interface Science. 77 (1), 91-101 (1980).
  102. Muller, V. M., Yushchenko, V. S., Derjaguin, B. V. General theoretical consideration of the influence of surface forces on contact deformations and the reciprocal adhesion of elastic spherical particles. Journal of Colloid and Interface Science. 92 (1), 92-101 (1983).
  103. Johnson, K. L., Greenwood, J. A. An adhesion map for the contact of elastic spheres. Journal of Colloid and Interface Science. 192 (2), 326-333 (1997).
  104. Shi, X., Zhao, Y. -. P. Comparison of various adhesion contact theories and the influence of dimensionless load parameter. Journal of Adhesion Science and Technology. 18 (1), 55-68 (2004).

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Atomic Force MicroscopyAFMCantilever-based NanoindentationNanoscaleMechanical PropertiesModulusAirFluidTopographical ImagingQuantitative MeasurementsCalibrationProbeForceDisplacementDeflection Sensitivity
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Enrriques, A. E., Howard, S., Timsina, R., Khadka, N. K., Hoover, A. N., Ray, A. E., Ding, L., Onwumelu, C., Nordeng, S., Mainali, L., Uzer, G., Davis, P. H. Atomic Force Microscopy Cantilever-Based Nanoindentation: Mechanical Property Measurements at the Nanoscale in Air and Fluid. J. Vis. Exp. (190), e64497, doi:10.3791/64497 (2022).
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