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Laser Capture Microdissection

Methods and Protocols

Series: Methods in Molecular Biology

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by Graeme I. Murray and Stephanie Curran

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Main description:

Expert laboratorians noted for their optimized methods describe in step-by-step detail how to successfully carry out tissue-based laser microdissection in their own laboratory using the different systems available and to apply a wide range of sophisticated molecular technologies to the microdissected tissue samples. Highlights include gene expression profiling of primary tumor cell populations using laser capture microdissection, RNA transcript amplification and GeneChip® microarrays, laser capture microdissection and colorectal cancer proteomics, and proteomic analysis of human bladder tissue using SELDI® approach following microdissection techniques. Applications of the microdissected tissue described encompass PCR, gene expression studies including microarray analysis, and proteomics.

Back cover:

Use of laser microdissection has grown enormously to become a very powerful high-throughput technology essential to a range of downstream applications in pathology, cancer research, life sciences, medical diagnostics, and biotechnology. In Laser Capture Microdissection, expert laboratory researchers noted for their optimized methods describe in step-by-step detail not only how to successfully carry out tissue-based laser microdissection in their own laboratories using the different systems available, but also how to apply a wide range of sophisticated molecular technologies to the microdissected tissue samples. These readily reproducible methods make it possible to carry out contamination-free molecular analyses of specific types of normal and diseased cells. Highlights include gene expression profiling of primary tumor cell populations, RNA transcript amplification and GeneChip® microarrays, colorectal cancer proteomics, and proteomic analysis of human bladder tissue using the SELDI® approach. Applications of the microdissected tissue described encompass PCR, gene expression studies including microarray analysis, and proteomics. The protocols follow the successful Methods in Molecular Biology™ series format, each offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.

Comprehensive and state-of-the-art, Laser Capture Microdissection provides a highly practical guide to successful tissue-based laser microdissection using available commercial systems and to the interpretation of the results using a wide range of sophisticated molecular technologies.

Contents:

Part I. Introduction

An Introduction to Laser-Based Tissue Microdissection Techniques
Stephanie Curran and Graeme I. Murray

Part II. Microdissection and DNA Analysis

Methacarn Fixation for Genomic DNA Analysis in Microdissected Cells
Makoto Shibutani and Chikako Uneyama

Multiplex Quantitative Real-Time PCR of Laser Microdissected Tissue
Patrick H. Rooney

Comparative Genomic Hybridization Using DNA From Laser Capture Microdissected Tissue
Grace Callagy, Lucy Jackson, and Carlos Caldas

Detection of Ki-ras and p53 Mutations by Laser Capture Microdissection/PCR/SSCP
Deborah Dillon, Karl Zheng, Brina Negin, and José Costa

Whole-Genome Allelotyping Using Laser Microdissected Tissue
Colleen M. Feltmate and Samuel C. Mok

Microdissection for Detecting Genetic Aberrations in Early and Advanced Human Urinary Bladder Cancer
Arndt Hartmann, Robert Stoehr, Peter J. Wild, Wolfgang Dietmaier, and Ruth Knuechel

Laser Microdissection for Microsatellite Analysis in Colon and Breast Cancer
Peter J. Wild, Robert Stoehr, Ruth Knuechel, Arndt Hartmann, and Wolfgang Dietmaier

Assessment of RET/PTC Oncogene Activation in Thyroid Nodules Utilizing Laser Microdissection Followed by Nested RT-PCR
Giovanni Tallini and Guilherme Brandao

Combined Laser-Assisted Microdissection and Short Tandem Repeat Analysis for Detection of In Situ Microchimerism After Solid Organ Transplantation
Ulrich Lehmann, Anne Versmold, and Hans Kreipe

Part III. RNA and Gene Expression

Studies Using Microdissected Cells, Laser-Assisted Microdissection of Membrane-Mounted Tissue Sections
Lise Mette Gjerdrum and Stephen Hamilton-Dutoit.

Laser-Assisted Microdissection of Membrane-Mounted Sections Following Immunohistochemistry and In Situ Hybridization
Lise Mette Gjerdrum and StephenHamilton-Dutoit

Laser-Assisted Cell Microdissection Using the PALM System
Patrick Micke, Arne Östman, Joakim Lundeberg, and Fredrik Ponten

Laser Microdissection and RNA Analysis
Ludger Fink and Rainer Maria Bohle

Gene Expression Profiling of Primary Tumor Cell Populations Using Laser Capture Microdissection, RNA Transcript Amplification, and GeneChip® Microarrays
Veronica I. Luzzi, Victoria Holtschlag, and Mark A. Watson

Quantification of Gene Expression in Mouse and Human Renal Proximal Tubules
Jun-ya Kaimori, Masaru Takenaka, and Kousaku Okubo

Laser Capture Microdissection for Analysis of Macrophage Gene Expression From Atherosclerotic Lesions
Eugene Trogan and Edward A. Fisher

Analysis of Pituitary Cells by Laser Capture Microdissection
Ricardo V. Lloyd, Xiang Qian, Long Jin, Katharina Ruebel, Jill Bayliss, Shuya Zhang, and Ikuo Kobayaski

Part IV. Microdissection Techniques and Applications in Proteomics

Laser Capture Microdissection and Colorectal Cancer Proteomics
Laura C. Lawrie and Stephanie Curran

Proteomic Analysis of Human Bladder Tissue Using SELDI® Approach Following Microdissection Techniques
Rene C. Krieg, Nadine T. Gaisa, Cloud P. Paweletz, and Ruth Knuechel

Part V. Microdissection and Molecular Analysis of Microorganisms

Genetic Analysis of HIV by In Situ PCR-Directed Laser Capture Microscopy of Infected Cells
Daniele Marras

Use of Laser Capture Microdissection Together With In Situ Hybridization and Real-Time PCR to Study the Distribution of Latent Herpes Simplex Virus Genomes in Mouse Trigeminal Ganglion
Xiao-Ping Chen, Marina Mata, and David J. Fink

Laser Capture Microdissection and PCR for Analysis of Human Papilloma Virus Infection
Kheng Chew, Patrick H. Rooney, Margaret E. Cruickshank, and Graeme I. Murray

Laser Capture Microdissection

PRODUCT DETAILS

ISBN-13: 9781617374586
Publisher: Springer (Humana Press)
Publication date: December, 2010
Pages: 320
Weight: 448g
Availability: Not available (reason unspecified)
Subcategories: Biochemistry

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From the reviews:

"...an excellent help to researchers...." - Neoplasma

"Comprehensive and state of the art, this book provides a highly practical guide to successful tissue-based laser microdissection using available commercial systems, and also a guide to interpreting the results using a wide range of sophisticated molecular technologies." - Clinical Laboratory International

"...provides researchers with practical advise and detailed technical protocols for carrying out tissue-based LCM...." - Vet Pathol

"Comprehensive and up-to-date collection of readily reproducible preparative and analytical methods is presented in each chapter for an in-depth practice of laser microdissection application. … The book is subdivided into 5 parts with a total of 24 chapters featuring step-by-step protocols that will enable novice researchers to isolate specific cell types or microorganisms." (Carlo Alberto Redi, European Journal of Histochemistry, December, 2005)