This new volume, number 123, of Methods in Cell Biology looks at methods for quantitative imaging in cell biology. It covers both theoretical and practical aspects of using optical fluorescence microscopy and image analysis techniques for quantitative applications.
The introductory chapters cover fundamental concepts and techniques important for obtaining accurate and precise quantitative data from imaging systems. These chapters address how choice of microscope, fluorophores, and digital detector impact the quality of quantitative data, and include step-by-step protocols for capturing and analyzing quantitative images. Common quantitative applications, including co-localization, ratiometric imaging, and counting molecules, are covered in detail. Practical chapters cover topics critical to getting the most out of your imaging system, from microscope maintenance to creating standardized samples for measuring resolution. Later chapters cover recent advances in quantitative imaging techniques, including super-resolution and light sheet microscopy. With cutting-edge material, this comprehensive collection is intended to guide researchers for years to come.
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Table of Contents
1. Concepts in Quantitative Fluorescence Microscopy2. Practical Considerations of Objective Lenses for Application in Cell Biology
3. Assessing Camera Performance for Quantitative Microscopy
4. A Practical Guide to Microscope Care and Maintenance
5. Fluorescence Live Cell Imaging
6. Fluorescent Proteins for Quantitative Microscopy: Important Properties and Practical Evaluation
7. Quantitative Confocal Microscopy: Beyond a Pretty Picture
8. Assessing and Benchmarking Multiphoton Microscopes for Biologists
9. Spinning Disk Confocal Microscopy: Present Technology and Future Trends
10. Quantitative Deconvolution Microscopy
11. Light Sheet Microscopy
12. DNA Curtains: Novel Tools for Imaging Protein-nucleic Acid Interactions at the Single-molecule Level
13. Nanoscale Cellular Imaging with Scanning Angle Interference Microscopy
14. Localization Microscopy in Yeast
15. Imaging Cellular Ultrastructure by PALM, iPALM, and Correlative iPALM-EM
16. Seeing more with Structured Illumination Microscopy
17. Structured Illumination Super-Resolution Imaging of the Cytoskeleton
18. Analysis of Focal Adhesion Turnover: A Quantitative Live Cell Imaging Example
19. Determining Absolute Protein Numbers by Quantitative Fluorescence Microscopy
20. High Resolution Traction Force Microscopy
21. Experimenters' Guide to Co-Localization Studies: Finding a Way Through Indicators and Quantifiers, in Practice
22. User-Friendly Tools for Quantifying the Dynamics of Cellular Morphology and Intracellular Protein Clusters
23. Ratiometric Imaging of pH Probes
24. Towards Quantitative Fluorescence Microscopy with DNA Origami Nanorulers
25. Imaging and Physically Probing Kinetochores in Live Dividing Cells
26. Adaptive Fluorescence Microscopy by Online Feedback Image Analysis
27. Open Source Solutions for SPIMage Processing
28. Second Harmonic Generation Imaging of Cancer
