Regular price
70.250 KD
inc. VAT
Couldn't load pickup availability
Table of contents
- Contentsv
- Contributorsxiii
- Prefacexix
- Part I: Basic Concept and Preparation Culture Substrates for Cell Mechanical Studies1
- Chapter 1: Basic Rheology for Biologists3
- I. Introduction and Rationale4
- II. Rheological Concepts6
- A. Elasticity7
- B. Viscosity8
- C. Oscillatory Measurements9
- III. Rheological Instrumentation11
- IV. Experimental Design13
- A. Stress-Strain Relation13
- B. Stress Relaxation14
- C. Creep and Creep Recovery15
- D. Frequency Sweep16
- E. Time Sweep17
- F. Strain or Stress Amplitude Sweeps18
- G. Rate-Dependent Viscosity19
- H. Flow Oscillation20
- V. Sample Preparation20
- A. Solids20
- B. Liquids and Gelling Systems22
- VI. Special Considerations for Biological Samples22
- A. Biological Polymers22
- B. Intact Tissue23
- C. Instrument Selection for Measuring Gelation Kinetics23
- VII. Conclusions24
- Glossary25
- References26
- Chapter 2: Polyacrylamide Hydrogels for Cell Mechanics: Steps Toward Optimization and Alternative Us29
- I. Introduction30
- II. Principle of the Polyacrylamide Hydrogel31
- A. Standard Method of Polymerization31
- B. Light-Induced Initiation of Polymerization31
- III. Conjugation of Proteins to Polyacrylamide33
- A. Carbodiimide-Mediated Cross-Linking34
- B. Activation with Acrylic Acid N-Hydroxysuccinimide Ester36
- C. Activation with the N-Succinimidyl Ester of Acrylamidohexanoic Acid (N6)38
- D. Other Protein-Coupling Methods40
- IV. Optimizing the Placement of Beads for Traction Force Microscopy40
- V. Manipulation of Gel Geometry41
- A. Preparation of Polyacrylamide Microbeads41
- B. Preparation of a Model Three-Dimensional Culture System42
- VI. Concluding Remarks44
- References45
- Chapter 3: Microscopic Methods for Measuring the Elasticity of Gel Substrates for Cell Culture: Micr47
- I. Introduction48
- II. Probing with Microspheres Under Gravitational Forces49
- III. Atomic Force Microscopy50
- IV. Probing with Spherically Tipped Glass Microindenters53
- A. Preparation and Calibration of the Spherically Tipped Microindenter54
- B. Calibration of the Microscope and Micromanipulator54
- C. Characterization and Calibration of the Microindenter55
- D. Measurement of the Indentations of Hydrogels in Response to Forces of the Microindenter57
- E. Data Analysis61
- F. Discussion62
- V. Conclusions64
- References64
- Chapter 4: Surface Patterning67
- I. Introduction68
- II. Patterning with Electrodynamic Instabilities69
- A. Procedure for Patterning with Electrohydrodynamic Instabilities71
- B. Cell Migration on Topographic Surfaces72
- III. Lithography Without a Clean Room73
- A. Photolithography Basics74
- IV. Patterning at the Micro- and Nanoscale with Polymer Mixtures and Block Copolymers80
- A. Principles and Procedures for Controlling Pattern Formation with Polymer Mixtures and Block Copol81
- V. Summary84
- Acknowledgments84
- References85
- Chapter 5: Molecular Engineering of Cellular Environments: Cell Adhesion to Nano-Digital Surfaces89
- I. Introduction: Sensing Cellular Environments90
- II. Nano-Digital Chemical Surfaces for Regulating Transmembrane-Receptor Clustering95
- A. Extended Nanopatterns and Biofunctionalization95
- B. Micro-Nanopatterns for Spatially Controlled Molecular Clustering97
- C. Cellular Responses to Nano-Digital and Biofunctionalized Surfaces101
- D. Local Versus Global Effects of Ligand Density on Cell Adhesion104
- E. Cell Spreading and Migration on Different Nanopatterns106
- F. High-Resolution Visualization of Cells in Contact with Biofunctionalized Nanopatterns107
- III. Outlook for the Future108
- Acknowledgments108
- References109
- Part II: Subcellular Mechanical Properties and Activities113
- Chapter 6: Probing Cellular Mechanical Responses to Stimuli Using Ballistic Intracellular Nanorheolo115
- I. Introduction117
- A. Why Cell Mechanics?117
- B. Particle-Tracking Nanorheology: Measuring the Local Viscoelastic Properties of a Cell by Tracking118
- C. Local and Global Viscoelastic Properties of the Cell119
- D. Interstitial Versus Mesoscale Viscosity of the Cytoplasm119
- E. Viscoelastic Properties of the Cytoplasm Depend on the Timescale of Applied Forces120
- F. The Viscoelastic Properties of the Cytoplasm Depend on the Amplitude of Applied Forces121
- G. Measurements at the Cell Surface Versus in the Cytoplasm121
- H. Methods of Delivery of Nanoparticles to the Cytoplasm123
- I. BIN: Proof of Principle124
- J. Intracellular Micromechanics of Cells in 3D Matrix125
- II. Materials and Instrumentation127
- A. Preparation of Nanoparticles127
- B. Cell Culture127
- C. Ballistic Injection of Nanoparticles129
- D. Encapsulation of Cells in a 3D Matrix129
- E. Imaging of Fluctuating Nanoparticles Embedded in the Cell129
- III. Procedures129
- A. Preparation of Nanoparticles129
- B. Ballistic Injection of Nanoparticles130
- C. Cell Seeding and Encapsulation in a 3D Matrix131
- D. BIN Analysis132
- IV. Pearls and Pitfalls135
- V. Concluding Remarks136
- A. Unique Advantages of BIN136
- B. Advantages of Traditional Particle-Tracking Nanorheology Are Maintained by BIN136
- Acknowledgments137
- References137
- Chapter 7: Multiple-Particle Tracking and Two-Point Microrheology in Cells141
- I. Introduction142
- II. Principles of Passive Tracer Microrheology146
- A. Conventional Passive Microrheology146
- B. Expected Tracer Motion and Tracking Performance147
- C. Two-Point Microrheology148
- III. Multiple-Particle Tracking Algorithms149
- A. Image Restoration150
- B. Locating Possible Particle Positions150
- C. Eliminating Spurious or Unwanted Particle Trajectories151
- D. Linking Positions into Trajectories152
- E. Available Software Packages and Computing Resources154
- IV. Computing Rheology from Tracer Trajectories155
- A. Computing Mean-Squared Displacements155
- B. Computing Two-Point Mean-Squared Displacements157
- C. Applying Automated Image Analysis for Statistics158
- D. Converting MSDs to Rheology159
- V. Error Sources in Multiple-Particle Tracking161
- A. Random Error (Camera Noise)162
- B. Systematic Errors163
- C. Dynamic Error165
- D. Sample Drift, Computational Detrending, and Its Limitations166
- E. Effects of Measurement Errors on the MSD167
- VI. Instrument Requirements for High-Performance Tracking168
- A. Isolation from Vibration, Acoustic Noise, and Thermal Drift168
- B. Microscopy-Generation of High-Contrast Tracer Images168
- C. Using Low-Noise, Non-interlaced Camera170
- D. Using High-Intensity, Filtered Illuminator171
- E. Using Synthetic Tracers to Increase Visibility172
- VII. Example: Cultured Epithelial Cells172
- A. Particle-Tracking Results for TC7 Epithelial Cells172
- B. TPM of TC7 Epithelial Cells174
- C. Computing Stress Fluctuation Spectra176
- VIII. Conclusions and Future Directions177
- References177
- Chapter 8: Imaging Stress Propagation in the Cytoplasm of a Living Cell179
- I. Introduction180
- II. Detecting External Stress-Induced Displacements in the Cytoplasm181
- A. Green Fluorescent Protein Transfection and Cell Culture181
- B. Application of Periodic Mechanical Stress182
- C. Image Acquisition182
- D. Image Partitioning183
- E. Array Shifting184
- F. Image Match Searching185
- G. Synchronous Displacement185
- H. Signal-to-Noise Ratio187
- III. Imaging Displacement and Stress Maps in a Live Cell188
- A. Quantifying Displacement Maps188
- B. Computing Stress Maps from Displacement Maps190
- C. Modulation of Stress Distribution Within a Living Cell190
- D. Application of Mechanical Loads in Any Direction Using 3D-MTC192
- E. Mechanical Anisotropic Signaling to the Cytoskeleton and to the Nucleolus195
- F. Limitations of 3D-MTC197
- IV. Future Prospects197
- Acknowledgments197
- References198
- Chapter 9: Probing Intracellular Force Distributions by High-Resolution Live Cell Imaging and Invers199
- I. Introduction200
- II. Methods202
- A. Actin Cytoskeleton Mechanics in Cell Protrusion202
- B. Force Reconstruction212
- C. Probing Heterogeneous Network Elasticity with Speckle Microscopy221
- III. Summary227
- IV. Appendix228
- A. Solution of the Inverse Problem228
- Acknowledgments231
- References231
- Chapter 10: Analysis of Microtubule Curvature237
- I. Introduction238
- II. Rationale240
- III. Raw Data Collection242
- A. Point-Click Method243
- B. Semiautomated Methods243
- C. Data Collection Errors244
- IV. Validation Strategy245
- A. Modeling of Semiflexible Polymers247
- B. Generation of Simulated Data248
- C. Validation of Semiflexible Polymer Simulation253
- V. Curvature Estimation Methods256
- A. Three-Point Method256
- B. Shape-Fitting Method256
- C. Constructing the Curvature Distribution257
- VI. Results258
- A. Three-Point Method258
- B. Shape-Fitting Method262
- VII. Discussion264
- VIII. Conclusions265
- Acknowledgments265
- References266
- Chapter 11: Nuclear Mechanics and Methods269
- I. Introduction270
- II. Experimental Methods for Probing Nuclear Mechanical Properties273
- A. Isolation of Individual Nuclei273
- B. Micropipette Aspiration Experiments275
- C. Substrate Strain Experiments277
- D. Compression Experiments280
- E. Indentation by AFM285
- F. Particle-Tracking Microrheology286
- G. Microneedle-Imposed Extension287
- III. Discussion and Prospects288
- IV. Outlook290
- References291
- Part III: Cellular and Embryonic Mechanical Properties and Activities295
- Chapter 12: The Use of Gelatin Substrates for Traction Force Microscopy in Rapidly Moving Cells297
- I. Introduction298
- II. Rationale299
- III. Methods300
- A. Preparation of Gelatin Substrates300
- B. Plating Cells onto a Gelatin Substrate302
- C. Data Collection and Analysis303
- D. Trouble-Shooting Guide305
- IV. Applications of the Gelatin Traction Force Assay to Study Mechano-signal Transduction in Moving307
- V. Other Applications and Future Directions309
- VI. Summary310
- Acknowledgments310
- References310
- Chapter 13: Microfabricated Silicone Elastomeric Post Arrays for Measuring Traction Forces of Adhere313
- I. Introduction314
- II. Microfabrication of the Micropost Arrays316
- A. Standard Photolithography317
- B. SU-8 Photolithography for Micropost Arrays318
- C. Soft Lithography320
- D. Troubleshooting anf Helpful Suggestions321
- III. Characterization of Micropost Spring Constant321
- A. Beam-Bending Theory321
- B. Measurement of Micropost Stiffness322
- IV. Analysis of Traction Forces Through Micropost Deflections323
- A. Substrate Preparation323
- B. Staining and Microscopy of Micropost Arrays325
- C. Image Analysis Techniques325
- V. Experimental Applications of Microposts and Discussion326
- Acknowledgments327
- References327
- Chapter 14: Cell Adhesion Strengthening: Measurement and Analysis329
- I. Introduction330
- II. The Cell Adhesion Process330
- III. Measurement Systems for Adhesion Characterization331
- IV. Hydrodynamic Assay for Quantifying Adhesion Strength334
- A. Experimental Design334
- B. Interpretation of Adhesion Strength Results337
- V. Quantitative Biochemical Methods for Adhesion Analysis338
- A. Quantification of Bound Integrin338
- B. Wet-Cleaving. Assay for Localized FA Protein Quantification338
- C. Immunofluorescence Staining and Quantification339
- VI. Simple Mathematical Modeling of Adhesion Strengthening Mechanics341
- A. Resolving Forces for a Cell Under Hydrodynamic Shear341
- B. Mathematical Analysis of Adhesion Strengthening Mechanics343
- VII. Discussion344
- Acknowledgments344
- References344
- Chapter 15: Studying the Mechanics of Cellular Processes by Atomic Force Microscopy347
- I. Introduction348
- II. Instrumentation and Operation Modes349
- A. Principal Components349
- B. Cantilevers350
- C. Combination of Optical Microscopy and AFM352
- III. Operating Modes353
- A. Imaging Modes353
- B. Force Curve Mode357
- C. Force Volume Mode358
- IV. Investigations of Live Cells358
- A. Imaging358
- B. Measuring Stiffness358
- C. Dynamics of Cellular Mechanical Properties365
- D. Effects of Cell Stiffness on AFM Imaging367
- V. Outlook368
- Acknowledgments369
- References369
- Chapter 16: Using Force to Probe Single-Molecule Receptor-Cytoskeletal Anchoring Beneath the Surface373
- I. Generic Methods and Physical Foundations374
- A. Using Force to Probe Single-Molecule Interactions374
- B. Testing Bonds on Solid Substrates375
- C. Simple Physics of Breaking a Bond377
- D. Forcing Bonds to Dissociate Faster Than Their Spontaneous Off Rate379
- E. Force Ramp Method and Examples of Testing Protein-Protein Interactions In Vitro380
- II. Probing Bonds at Cell Surfaces383
- A. Phenomenology of Unbinding Events383
- B. Experimental Frustrations386
- C. Most Likely Site of Molecular Unbinding When Probing Bonds at Cell Surfaces387
- D. Adhesive Failure Without Membrane-Cytoskeletal Separation388
- E. Membrane-Cytoskeletal Unbinding Followed by Adhesive Detachment390
- F. Overlapping Bond Failure Processes391
- III. Future Challenge and Opportunity393
- Acknowledgments395
- References395
- Chapter 17: High-Throughput Rheological Measurements with an Optical Stretcher397
- I. Introduction398
- II. Rationale401
- III. Methods402
- A. Basic Experimental Setup402
- B. Preparation of Cells404
- C. Measurement Process404
- D. Computer Control of the Measurement405
- E. Image Analysis406
- F. Interpretation of the Data407
- IV. Additional Notes on Equipment412
- A. Optical Fibers412
- B. Laser Source413
- C. Microfluidic Chip414
- D. Microscopy Technique418
- E. Camera419
- V. Discussion419
- VI. Summary421
- Acknowledgments421
- References421
- Chapter 18: Measuring Mechanical Properties of Embryos and Embryonic Tissues425
- I. Introduction426
- A. Biomechanics and Developmental Biology426
- B. Why Do We Want to Understand the Mechanical Basis of Morphogenesis?427
- II. Applying and Measuring Forces of 10 nN to 10 muN428
- III. Nanonewton Force Apparatus: Parts, Function, and Operation431
- IV. Preparation of Tissue Samples432
- V. Measurement of the Time-Dependent Elasticity of Embryos or Tissue Explants434
- VI. Spring and Dashpot Models of Viscoelasticity Represent More Complex Structural Sources435
- VII. Challenges of Working with Embryonic Tissues435
- VIII. Use of Standard Engineering Terms and Units437
- IX. Future Prospects437
- References437
- Part IV: Mechanical Stimuli to Cells441
- Chapter 19: Tools to Study Cell Mechanics and Mechanotransduction443
- I. Introduction444
- II. Control of Cell Shape, Cytoskeletal Organization, and Cell Fate Switching446
- A. Microcontact Printing of Micropatterned Substrates for Cell Culture447
- B. Application Notes on Microcontact Printing449
- C. Extension and Future Development of Microcontact Printing453
- III. Probing Cell Mechanics, Cytoskeletal Structure, and Mechanotransduction454
- A. Magnetic Twisting Cytometry (MTC)455
- B. Applications of MTC458
- C. Extension and Future Development of MTC459
- D. Magnetic Pulling Cytometry (MPC)460
- E. Applications of MPC467
- IV. Discussion and Future Implications467
- References469
- Chapter 20: Magnetic Tweezers in Cell Biology473
- I. Introduction474
- II. Physics of Magnetic Tweezers475
- III. Magnetic Field Considerations477
- A. Sources of Magnetic Field477
- B. Magnet Pole Design478
- IV. Magnetic Particle Selection479
- V. Basic Solenoid Apparatus481
- VI. Force Calibration482
- A. Calibration Sample Protocol484
- B. Calibration Procedure485
- C. Direction of Magnetic Force485
- D. Bread-Tracking System486
- E. Data Interpretation486
- VII. Experimental Procedures487
- References491
- Chapter 21: Optical Neuronal Guidance495
- I. Introduction496
- A. Neuron Structure496
- B. Neuronal Cells in Development496
- C. Growth Cone Movement and Guidance498
- D. Existing Guidance Methods500
- II. Apparatus501
- A. Laser Light Sources501
- B. Laser Light Control Elements502
- C. Microscope Irradiation and Imaging504
- D. Cell Culture System504
- E. Adapting Existing Systems for Optical Guidance508
- III. Experiments509
- A. Turns509
- B. Accelerated Growth510
- C. Bifurcations511
- D. Other Observations511
- IV. Plausible Mechanisms of Optical Guidance513
- A. Filopodial Asymmetries513
- B. Retrograde Flow515
- C. Actin Polymerization via Membrane Tweezing515
- D. Laser-Induced Heating515
- V. Summary516
- Acknowledgments517
- References517
- Chapter 22: Microtissue Elasticity: Measurements by Atomic Force Microscopy and Its Influence on Cel521
- I. Introduction522
- II. AFM in Microelasticity Measurements526
- A. AFM Probing and Analysis526
- B. General Issues in Sample Preparation530
- III. Materials Characterization531
- A. Artificial Matrices531
- B. Cell-Secreted Matrices536
- C. Passive Tissue Elasticity538
- IV. Assessing Mechanical Influences on Cells541
- References542
- Chapter 23: Demystifying the Effects of a Three-Dimensional Microenvironment in Tissue Morphogenesis547
- I. Introduction548
- II. Rationale550
- A. Stromal-Epithelial Interactions550
- B. ECM Mechanics and Epithelial Behavior551
- C. 3D Organotypic Model Systems556
- III. Methods558
- A. Engineered Cell/Tissue Explants558
- B. Isolation of Bulk Proteins565
- C. Isolation of Bulk mRNA567
- D. Rapid Protein Isolation Techniques569
- E. Immunofluorescence571
- IV. Materials573
- A. Engineering Tissue Explants573
- B. Isolation of Bulk Proteins575
- C. Isolation of Bulk mRNA575
- D. Rapid Protein Isolation Techniques576
- E. Immunofluorescence576
- V. Discussion577
- Acknowledgments579
- References580
- Index585
- Volumes in Series601
Book details
- Vendor Elsevier S & T
- SKU 9780123705006
- ISBN-13 9780080548708
- Author Wang, Yu-Li; Discher, Dennis E.
- Category Science
- Subject Biotechnology
Do you have questions about this book?
Cell mechanics is the field of study that looks at how cells detect, modify, and respond to the physical properties of the cell environment. Cells communicate with each other through chemical and physical signals which are involved in a range of process from embryogenesis and wound healing to pathological conditions such as cancerous invasion. Similar principles are also likely to be critical for success in regenerative medicine. Cell mechanics is thus central to understanding these principles. As cell mechanics draws from the fields of biology, chemistry, physics, engineering, and mathematics, this book aims not only to provide a collection of research methods, but also to develop a common language among scientists who share the interest in cell mechanics but enter the field with diverse backgrounds. To this end all of the contributing authors have sought to explain in plain language the nature of the biological problems, the rationale for the approaches, in addition to the methods themselves. In addition, to balance practical utility against conceptual advances, the book has intentionally included both chapters that provide detailed recipes and those that emphasize basic principles.
* Presents a distinctive emphasis on matrix mechanics and their interplay with cell functions
* Includes highly significant topics relevant to basic and translational research, as well as tissue engineering
* Emphasizes mechanical input and output of cells
* Presents a distinctive emphasis on matrix mechanics and their interplay with cell functions
* Includes highly significant topics relevant to basic and translational research, as well as tissue engineering
* Emphasizes mechanical input and output of cells
Instant delivery by email
Your access email arrives within minutes of checkout, with a sign-in link for each book — no shipping, no waiting.
Read on any device
Books open in VitalSource Bookshelf on your phone, tablet, or computer, online or offline. Your library is always available at aafaq.vitalsource.com — just log in with the email you used at checkout.
Lost the email?
Resend it to yourself in seconds from My eBook orders, or email cs@aafaqeducation.com and we'll help.