Investigations of Cellular and Molecular Biophysical Properties by Atomic Force Microscopy Nanorobotics (eBook)

Dr. Mi Li received his B.E. and M.E. degree from Huazhong University of Science and Technology, Wuhan, China in 2006 and 2008, respectively, and Ph. D. degree from Shenyang Institute of Automation, Chinese Academy of Sciences, Shenyang, China in 2015. Currently Dr. Li is Associate Professor at the Shenyang Institute of Automation and his research areas include micro/nano automation technology, atomic force microscopy, nanorobot and cell mechanics. He has published more than 30 articles in academic journals as first author, and received Outstanding Doctoral Dissertation Award of Chinese Academy of Sciences in 2016. 

Supervisor’s Foreword 6
Parts of this thesis have been published in the following journal articles: 8
Acknowledgements 10
Contents 12
1 Introduction to Atomic Force Microscopy-Based Nanorobotics for Biomedical Applications 15
1.1 Background and Motivation 15
1.2 Cellular Physiological Properties Detection 18
1.3 Single-Cell and Single-Molecule Techniques 19
1.4 Nanorobot Based on AFM 24
1.5 Opportunities and Challenges 26
1.6 Thesis Contents and Chapter Organization 28
References 30
2 Immobilization Methods for Observing Living Mammalian Suspended Cells by AFM 35
2.1 Current Status of Immobilizing Living Cell for AFM Imaging 35
2.2 Biological Experiment Platform Based on AFM Nanorobot 36
2.3 Immobilizing Mammalian Suspended Cells by Micropillar Array 38
2.4 Imaging Living Mammalian Suspended Cells Based on Micro-Pillar Immobilization 40
2.5 Imaging Living Mammalian Suspended Cells Based on Micro-Well Immobilization 41
2.6 Summary 43
References 44
3 Measuring the Mechanical Properties of Single Cells by AFM 46
3.1 Current Status of Measuring Cell Mechanics by AFM 47
3.2 Principle and Methods of Measuring Cell Mechanics by AFM 48
3.3 Fabrication of AFM Spherical Tip 52
3.4 Measuring the Mechanics of Cancerous Cells with Different Invasive Abilities 53
3.5 Summary 58
References 58
4 Single-Molecule Recognition and Force Measurements by AFM 61
4.1 Current Status of AFM Single-Molecule Force Measurements 62
4.2 Principle of AFM Single-Molecule Force Spectroscopy 63
4.3 Tip Functionalization and Verification 65
4.4 Measuring the CD20-Rituximab Interaction Forces 68
4.4.1 Purified CD20s on Mica 68
4.4.2 Native CD20s on Lymphoma Cell 71
4.5 Summary 73
References 74
5 Mapping Membrane Proteins on Cell Surface by AFM 77
5.1 Current Status of Mapping Membrane Proteins on Cell Surface by AFM 77
5.2 Peak Force Tapping AFM 78
5.3 Mapping Protein Distributions by PFT 79
5.3.1 Avidins on Mica 80
5.3.2 CD20s on Lymphoma Cell Surface 83
5.4 Summary 87
References 88
6 Applications of AFM Cellular and Molecular Biophysical Detection in Clinical Lymphoma Rituximab Treatment 90
6.1 Cellular Ultra-Microstructure and Mechanics Detection 90
6.1.1 PCD Mechanism 90
6.1.2 ADCC Mechanism 94
6.1.3 CDC Mechanism 102
6.2 Probing Target Proteins on Primary Cells from Clinical Lymphoma Patients 110
6.2.1 Detecting CD20-Rituximab Interactions on Patient Cancerous Cells 110
6.2.2 Detecting FcR-Rituximab Interactions on Patient Effector Cells 121
6.3 Comparison of AFM Detection Results and Clinical Therapeutic Outcomes 130
6.3.1 CD20-Rituximab Interaction on Patient Cancerous Cells and Clinical Efficacies 130
6.3.2 FcR-Rituximab Interaction Patient Effector Cells and Clinical Efficacies 131
6.4 Summary 136
References 136
7 Conclusion and Future Work 140
7.1 Conclusion 140
7.2 Future Work 141
References 143
Curriculum Vitae 144