This volume explores various approaches for enrichment, detection, isolation, and molecular profiling of circulating tumor cells (CTCs). Each chapter provides comprehensive descriptions and guidelines on how to perform innovative experiments in CTC research. Included are protocols for capture of CTCs via filtration and density gradient centrifugation; microfluidic and immunomagnetic separation of CTCs; detection of CTCs by immunocytochemistry, fluorescence in situ hybridization, and flow cytometry; assays designed for genomic characterization and functional analyses of CTCs, and many more. Written in the highly successful Methods in Molecular Biology series format, the chapters in this book include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.Cutting-edge and authoritative, Circulating Tumor Cells: Methods and Protocols is a valuable resource for laboratory researchers and clinicians who are interested in furthering their studies on CTCs.
Dedication... Acknowledgement... Foreword... Preface... Table of Contents... Contributing Authors... 1. Microfluidic Capture and Multiplex Immunofluorescence of Circulating Tumor Cells to Identify Cancer Origin Chian-Hui Lai and Ying-Chih Chang 2. Microfluidic Separation of Circulating Tumor Cells Based on Size and Deformability Emily S. Park, Simon P. Duffy, and Hongshen Ma 3. A Novel Microfluidic Device for Isolation of Circulating Tumor Cells from Pancreatic Cancer Blood Samples Jose I. Varillas, Kangfu Chen, Jinling Zhang, Thomas J. George, Jr., and Z. Hugh Fan 4. Microfluidic-Based Enrichment and Retrieval of Circulating Tumor Cells for RT-PCR Analysis Priya Gogoi, Saedeh Sepehri, Will Chow, Kalyan Handique, and Yixin Wang 5. Micro-Scale Laminar Vortices for High-Purity Extraction and Release of Circulating Tumor Cells Soojung Claire Hur, James Che, and Dino Di Carlo 6. Separable Bilayer Microfiltration Device for Viable Label-free Enrichment of Circulating Tumor Cells Sijie Hao, Merisa Nisic, Hongzhang He, Yu-Chong Tai, and Si-Yang Zheng 7. Microfilter-Based Capture and Release of Viable Circulating Tumor Cells Siddarth Rawal, Zheng Ao, Ram H. Datar, and Ashutosh Agarwal 8. Fourier Ptychographic Microscopy for Rapid, High Resolution Imaging of Circulating Tumor Cells Enriched by Microfiltration Anthony Williams, Jaebum Chung, Changhuei Yang, and Richard J. Cote 9. Automated Microfluidic Filtration and Immunocytochemistry Detection System for Capture and Enumeration of Circulating Tumor Cells and Other Rare Cell Populations in Blood Michael Pugia, Mark Jesus M. Magbanua, and John W. Park 10. Filter-Adapted Fluorescent In Situ Hybridization (FA-FISH) for Filtration-Enriched Circulating Tumor Cells Marianne Oulhen, Emma Pailler, Vincent Faugeroux, and Francoise Farace 11. Negative Enrichment and Isolation of Circulating Tumor Cells for Whole Genome Amplification Nisha Kanwar and Susan J. Done 12. Capture and Genetic Analysis of Circulating Tumor Cells Using a Magnetic Separation Device (Magnetic Sifter) Chin Chun Ooi, Seung-min Park, Dawson J. Wong, Sanjiv S. Gambhir, and Shan X. Wang 13. RareCyte (R) CTC Analysis Step 1: AccuCyte (R) Sample Preparation for the Comprehensive Recovery of Nucleated Cells from Whole Blood Arturo B. Ramirez, Lance U'Ren, Danny Campton, David Stewart, Joshua J. Nordberg, Jackie L. Stilwell, and Eric P. Kaldjian 14. RareCyte (R) CTC Analysis Step 2: Detection of Circulating Tumor Cells by CyteFinder (R) Automated Scanning and Semi-Automated Image Analysis Jeffrey L. Werbin, Joshua J. Nordberg, Jay Tzucker, Paulina Varshavskaya, Jackie L. Stilwell, and Eric P. Kaldjian 15. RareCyte (R) CTC Analysis Step 3: Using the CytePicker (R) Module for Individual Cell Retrieval and Subsequent Whole Genome Amplification of Circulating Tumor Cells for Genomic Analysis Jackie L. Stilwell, Paulina Varshavskaya, Jeffrey L. Werbin, Joshua J. Nordberg, Arturo B. Ramirez, Steve Quarre, Jay Tzucker, Jennifer Chow, Brennan Enright, and Eric P. Kaldjian 16. Enumeration, Dielectrophoretic Capture, and Molecular Analysis of Circulating Tumor Cells Stephanie S. Yee and Erica L. Carpenter 17. 18. Flow Cytometric Detection of Circulating Tumor Cells Using a Candidate Stem Cell Marker, p75 Neurotrophin Receptor (p75NTR)< 19. Multispectral Imaging Analysis of Circulating Tumor Cells in Negatively-Enriched Peripheral Blood Samples Brandon Miller, Maryam Lustberg, Thomas A. Summers, and Jeffrey J Chalmers 20. Fiber-Optic Array Scanning Technology (FAST) for Detection and Molecular Characterization of Circulating Tumor Cells Zheng Ao and Xiaohe Liu 21. A Non-Invasive and Real-Time Method for Circulating Tumor Cell Detection: In Vivo Flow Cytometry Xunbin Wei, Jian Zhou, Xi Zhu, Xinrong Yang, Ping Yang, and Qiyan Wang 22. EpCAM-Independent Enrichment and Detection of Viable CTCs Using the EPISPOT Assay Alexandra Soler, Laure Cayrefourcq, Martine Mazel, and Catherine Alix-Panabieres 23. Utilizing Matrigel Transwell Invasion Assay to Detect and Enumerate Circulating Tumor Cells Xingtong Liu and Xiangwei Wu 24. Circulating Tumor Cells: Markers and Methodologies for Enrichment and Detection M.J. Alvarez Cubero, J.A. Lorente, I. Robles-Fernandez, A. Rodriguez-Martinez, J.L. Puche, and M.J. Serrano