NanoWizard AFM Handbook - KTH The atomic force microscope (AFM) is one of the family of scanning probe

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  • NanoWizard® AFM Handbook Version 2.2a 05 / 2012

    © 2004-2012 JPK Instruments AG all rights reserved

  • JPK Instruments NanoWizard® Handbook Version 2.2a 1

    Contents

    1. Introduction ...........................................................................................................................................1 1.1 About this handbook ........................................................................................................................................ 1 1.2 What is an Atomic Force Microscope? ............................................................................................................ 1 1.3 Scanning Probe Microscopy ............................................................................................................................ 2 1.4 Atomic Force Microscopy ................................................................................................................................ 3 1.5 AFM cantilevers ............................................................................................................................................... 4

    2. Imaging modes .....................................................................................................................................5 2.1 Feedback and imaging control ......................................................................................................................... 5 2.2 Amplitude feedback in dynamic modes ........................................................................................................... 5 2.3 Another way of thinking about imaging modes ................................................................................................ 6 2.4 Operation ......................................................................................................................................................... 8 2.5 Phase imaging ................................................................................................................................................. 9 2.6 Force adjustment in imaging modes .............................................................................................................. 10 2.7 Applications ................................................................................................................................................... 11

    3. Force spectroscopy ............................................................................................................................12 3.1 Introduction .................................................................................................................................................... 12 3.2 Data processing for analysis .......................................................................................................................... 13 3.3 Applications ................................................................................................................................................... 16

    4. More about cantilevers .......................................................................................................................18 4.1 General points ............................................................................................................................................... 18 4.2 Handling information ...................................................................................................................................... 18 4.3 Cantilever types for different imaging modes ................................................................................................. 19 4.4 Tip modification ............................................................................................................................................. 20 4.5 Spring constant .............................................................................................................................................. 21

    5. Cell imaging ........................................................................................................................................24 5.1 AFM in relation to other cell imaging techniques ........................................................................................... 24 5.2 Sample preparation ....................................................................................................................................... 25 5.3 Imaging modes .............................................................................................................................................. 27 5.4 Critical imaging parameters ........................................................................................................................... 28 5.5 Using the oscilloscope to optimize the imaging parameters .......................................................................... 28 5.6 Artifacts .......................................................................................................................................................... 30

    6. Single molecule imaging .....................................................................................................................31 6.1 Preparation is key .......................................................................................................................................... 31 6.2 Imaging hints – intermittent contact mode in liquid ........................................................................................ 33 6.3 Imaging hints - contact mode in liquid ............................................................................................................ 34 6.4 Imaging hints – imaging in air ........................................................................................................................ 35 6.5 Simple DNA protocol for imaging tests .......................................................................................................... 35

    7. Artifacts ...............................................................................................................................................37 7.1 Tip shape issues ............................................................................................................................................ 37 7.2 Artifacts from damaged tips ........................................................................................................................... 38 7.3 Contamination ............................................................................................................................................... 39

  • JPK Instruments NanoWizard® Handbook Version 2.2

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    7.4 Other imaging considerations ........................................................................................................................ 39

    8. Useful physics for SPM ...................................................................................................................... 41 8.1 The cantilever resonance .............................................................................................................................. 41 8.2 Thermal noise spring constant calibration ..................................................................................................... 41 8.3 Young’s Modulus of materials ........................................................................................................................ 45

    9. Useful chemistry and sample/tip preparations ................................................................................... 47 9.1 Cleaning cantilevers and tips ......................................................................................................................... 47 9.2 Silanization and APTES treatment ................................................................................................................ 48 9.3 Home made gel packs for cantilever storage ................................................................................................. 49 9.4 Suppliers of AFM accessories ....................................................................................................................... 49

    10. References ......................................................................................................................................... 51 10.1 General AFM Papers ..................................................................................................................................... 51 10.2 Spring constant calibration references .......................................................................................................... 53 10.3 Books ............................................................................................................................................................ 54

  • JPK Instruments NanoWizard® Handbook Version 2.2a 1

    1. Introduction 1.1 About this handbook

    Here you can find information about the principles and methods of scanning probe microscopy. The focus is on applications in biotechnology and life science. The particular details of the JPK NanoWizard® AFM system, for both the software and hardware, can be found separately in the NanoWizard® User Manual. The aim of this document is to introduce AFM for those who are not familiar with the technique, and to provide background information and resources to aid those who are familiar with the technique to extend their knowledge of particular applications.

    The first sections of this handboo

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