Optimal molecular profiling of tissue and tissue components: De fining the best processing and microdissection methods for biomedical applications

Jaime Rodriguez-Canales, Jeffrey C. Hanson, Jason D. Hipp, Ulysses J. Balis, Michael A. Tangrea, Michael R. Emmert-Buck, G. Steven Bova

    Research output: Chapter in Book/Report/Conference proceedingChapterScientificpeer-review

    10 Citations (Scopus)

    Abstract

    Isolation of well-preserved pure cell populations is a prerequisite for sound studies of the molecular basis of any tissue-based biological phenomenon. This updated chapter reviews current methods for obtaining anatomically specific signals from molecules isolated from tissues, a basic requirement for productive linking of phenotype and genotype. The quality of samples isolated from tissue and used for molecular analysis is often glossed over or omitted from publications, making interpretation and replication of data difficult or impossible. Fortunately, recently developed techniques allow life scientists to better document and control the quality of samples used for a given assay, creating a foundation for improvement in this area. Tissue processing for molecular studies usually involves some or all of the following steps: tissue collection, gross dissection/identification, fixation, processing/embedding, storage/archiving, sectioning, staining, microdissection/annotation, and pure analyte labeling/identification and quantification. We provide a detailed comparison of some current tissue microdissection technologies and provide detailed example protocols for tissue component handling upstream and downstream from microdissection. We also discuss some of the physical and chemical issues related to optimal tissue processing and include methods specific to cytology specimens. We encourage each laboratory to use these as a starting point for optimization of their overall process of moving from collected tissue to high-quality, appropriately anatomically tagged scientific results. Improvement in this area will significantly increase life science quality and productivity. The chapter is divided into introduction, materials, protocols, and notes subheadings. Because many protocols are covered in each of these sections, information relating to a single protocol is not contiguous. To get the greatest benefit from this chapter, readers are advised to read through the entire chapter first, identify protocols appropriate to their laboratory for each step in their work flow, and then reread entries in each section pertaining to each of these single protocols.

    Original languageEnglish
    Title of host publicationPancreatic Cancer: Methods and Protocols
    PublisherHUMANA PRESS INC
    Pages61-120
    Number of pages60
    Volume980
    ISBN (Print)978-1-62703-286-5
    DOIs
    Publication statusPublished - 2013
    Publication typeA3 Book chapter

    Publication series

    NameMethods in Molecular Biology
    Volume980
    ISSN (Print)1064-3745

    Keywords

    • DNA
    • Fixation
    • Histopathology
    • Laser capture microdissection
    • Protein extraction
    • RNA
    • Sample preparation
    • laser microdissection
    • methods
    • molecular profiling
    • optimization

    Publication forum classification

    • Publication forum level 1

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