Laser Capture Microdissection Methods and Protocols

Laser microdissection techniques have revolutionized the ability of researchers in general, and pathologists in particular, to carry out molecular analysis on specific types of normal and diseased cells and to fully utilize the power of current molecular technologies including PCR, microarrays, and proteomics.  In second edition of Laser Capture Microdissection: Methods and Protocols, experts in the field provide the reader with practical advice on how to carry out tissue-based laser microdissection successfully in their own laboratory using the different laser microdissection systems that are available and to apply a wide range of molecular technologies.  The individual chapters encompass detailed descriptions of the individual laser based micro-dissection systems.  The downstream applications of the laser microdissected tissue described in the book include PCR in its many different forms as well as gene expression analysis including application to microarrays and proteomics. Written in the highly successful Methods in Molecular Biology™ series format, chapters 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.

1. Laser Capture Microdissection: Methods and Applications

            Kristen DeCarlo, Andrew Emley, Ophelia E. Dadzie, and Meera Mahalingam

2. Laser Microdissection for Gene Expression Profiling

            Lori A. Field, Brenda Deyarmin, Craig D. Shriver, Darrell L. Ellsworth, and Rachel E. Ellsworth

3. Gene Expression Using the PALM System

            Jian-Xin Lu and Cheuk-Chun Szeto

4. Immuno-Guided Microdissection Techniques

            Michael A. Tangrea, Jeffrey C. Hanson, Robert F. Bonner, Thomas J. Pohida, Jaime Rodriguez-Canales, and Michael R. Emmert-Buck

5. Optimized RNA Extraction from Non-Deparaffinized, Laser-Microdissected Material

            Danny Jonigk, Friedrich Modde, Clemens Bockmeyer, Jan Becker, and Ulrich Lehmann

6. Laser Capture Microdissection for Analysis of Gene Expression in Formalin-Fixed Paraffin-Embedded Tissue

            Ru Jiang, Rona S. Scott, and Lindsey M. Hutt-Fletcher

7. MicroRNA Profiling Using RNA from Microdissected Immunostained Tissue

            Clemens L. Bockmeyer, Danny Jonigk, Hans Kreipe, and Ulrich Lehmann

8. Profiling Solid Tumor Heterogeneity by LCM and Biological MS of Fresh Frozen Tissue Sections

            Donald J. Johann, Jr., Sumana Mukherjee, DaRue A. Prieto, Timothy D. Veenstra, and Josip Blonder

9. Amplification Testing in Breast Cancer by Multiplex Ligation-Dependent Probe Amplification of Microdissected Tissue

            Cathy B. Moelans, Roel A. de Weger, and Paul J. van Diest

10. Detection and Quantification of MicroRNAs in Laser Microdissected Formalin-Fixed Paraffin-Embedded Breast Cancer Tissues

            Sarkawt M. Khoshnaw, Des G. Powe, Ian O. Ellis, and Andrew R. Green

11. Laser Capture Microdissection Applications in Breast Cancer Proteomics

            René B.H. Braakman, Theo M. Luider, John W.M. Martens, John A. Foekens, and Arzu Umar

12. Proteomic Analysis of Laser Microdissected Ovarian Cancer Tissue with SELDI-TOF MS

            Isabelle Cadron, Toon Van Gorp, Philippe Moerman, Etienne Waelkens, and Ignace Vergote

13. LCM Assisted Biomarker Discovery from Archival Neoplastic Gastrointestinal Tissues

            Patricia A. Meitner and Murray B. Resnick

14. Purification of Diseased Cells from Barrett’s Esophagus and Related Lesions by Laser Capture Microdissection

            Masood A. Shammas and Manjula Y. Rao

15. Laser Microdissection of Intestinal Epithelial Cells and Downstream Analysis

            Benjamin Funke

16. Application of Laser Microdissection and Quantitative PCR to Assess the Response of Oesophageal Cancer to Neoadjuvant Chemo-Radiotherapy

            Claus Hann von Weyhern and Björn L.D.M. Brücher

17. Oligonucleotide Microarray Expression Profiling of Contrasting Invasive Phenotypes in Colorectal Cancer

            Christopher C. Thorn, Deborah Williams, and Thomas C. Freeman

18. Evaluation of Gastrointestinal mtDNA Depletion in Mitochondrial Neurogastrointestinal Encephalomyopathy (MNGIE)

            Carla Giordano and Giulia d’Amati

19. Laser Microdissection for Gene Expression Study of Hepatocellular Carcinomas Arising in Cirrhotic and Non-Cirrhotic Livers

            Maria Tretiakova and John Hart

20. Laser Capture Microdissection of Pancreatic Ductal Adeno-Carcinoma Cells to Analyze EzH2 by Western Blot Analysis

            Aamer M. Qazi, Sita Aggarwal, Donald W. Weaver, and Ramesh B. Batchu

21. Laser-Capture Microdissection of Renal Tubule Cells and Linear Amplification of RNA for Microarray Profiling and Real-Time PCR

            Susie-Jane Noppert, Susanne Eder, and Michael Rudnicki

22. Subcellular Renal Proximal Tubular Mitochondrial Toxicity with Tenofovir Treatment

            James J. Kohler and Seyed H. Hosseini

23. Application of Laser Capture Microdissection to Study Renal Carcinogenesis

            Kerstin Stemmer and Daniel R. Dietrich

24. Laser Capture Microdissection and Transcriptional Profiling in Archival FFPE Tissue in Prostate Cancer

            Ajay Joseph and Vincent J. Gnanapragasam

25. Quantitative Analysis of the Enzymes Associated with 5-Fluorouracil Metabolism in Prostate Cancer Biopsies

            Tomoaki Tanaka

26. Microdissection of Gonadal Tissues for Gene Expression Analyses

            Anne Jørgensen, Marlene Danner Dalgaard, and Si Brask Sonne

27. Duplex Real Time PCR Assay for Quantifying Mitochondrial DNA Deletions in Laser Microdissected Single Spiral Ganglion Cells

            Adam Markaryan, Erik G. Nelson, and Raul Hinojosa

28. Neuronal Type-Specific Gene Expression Profiling and Laser-Capture Microdissection

            Charmaine Y. Pietersen, Maribel P. Lim, Laurel Macey, Tsung-Ung W. Woo, and Kai C. Sonntag

29. Region-Specific In Situ Hybridization-Guided Laser Capture Microdissection on Postmortem Human Brain Tissue Coupled with Gene Expression Quantification

            René Bernard, Sharon Burke, and Ilan A. Kerman

30. UV-Laser Microdissection and mRNA Expression Analysis of Individual Neurons from Post Mortem Parkinson’s Disease Brains

            Jan Gründemann, Falk Schlaudraff, and Birgit Liss

31. Transcriptome Profiling of Murine Spinal Neurulation Using Laser Capture Microdissection and High-Density Oligonucleotide Microarrays

            Shoufeng Cao, Boon-Huat Bay, and George W. Yip

32. Probing the CNS Microvascular Endothelium by Immune-Guided Laser Capture Microdissection Coupled to Quantitative RT-PCR

            Nivetha Murugesan, Jennifer Macdonald, Shujun Ge, and Joel S. Pachter

33. Laser Capture Microdissection for Factor VIII-Expressing Endothelial Cells in Cancer Tissues

            Tomoatsu Kaneko, Takashi Okiji, Reika Kaneko, Hideaki Suda, and  Jacques E. Nör

34. Laser Capture Microdissection and Analysis of Liver Endothelial Cells from Patients with Budd Chiari Syndrome

            Selcuk Sozer and Ronald Hoffman

35. Laser Capture Microdissection of Hyperlipidemic/ApoE^-/- Mouse Aorta Atherosclerosis

            Michael Beer, Sandra Doepping, Markus Hildner, Gabriele Weber, Rolf Grabner, Desheng Hu, Sarajo Kumar Mohanta, Prasad Srikakulapu, Falk Weih, and Andreas J.R. Habenicht

36. Gene Expression Profiling in Laser Microdissected Bone Marrow Megakaryocytes

            Kais Hussein

37. Specific RNA Collection from the Rat Endolymphatic Sac by Laser Capture Microdissection (LCM): LCM of Very Small Organ Surrounded by Bony Tissue

            Kosuke Akiyama, Takenori Miyashita, Ai Matsubara, and Nozomu Mori

38. The Use of Laser Capture Microdissection on Adult Human Articular Cartilage for Gene Expression Analysis

            Naoshi Fukui, Yasuko Ikeda, and Nobuho Tanaka

39. Laser Capture Microdissection of Developing Barley Seeds and cDNA Array Analysis of Selected Tissues

            Johannes Thiel, Diana Weier, and Winfriede Weschke

40. Quantitative RT-PCR Gene Expression Analysis of a Laser Microdissected Placenta: An Approach to Study Preeclampsia

            Yuditiya Purwosunu, Akihiko Sekizawa, Takashi Okai, and Tetsuhiko Tachikawa