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This detailed volume examines fine-tuned methodologies using the planarian species, Schmidtea mediterranea. The book features experimental protocols covering topics from in situ hybridization, immunohistochemistry, cell dissociation and flow cytometry, to pipelines for the analysis of large datasets, as in genomics and transcriptomics. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Schmidtea mediterranea: Methods and Protocols provides both experts in the field and newcomers with the best possible toolbox for their everyday lab work utilizing this valuable model.
List of contents
Building Phylogenies from Transcriptomic Data.- Isolation and Library Preparation of Planarian piRNAs.- Genome-Wide Analysis of Planarian piRNAs.- Combining Fluorescent In Situ Hybridization with Immunofluorescence and Lectin Staining in Planarians.- Colorimetric Whole-Mount In Situ Hybridization in Planarians.- Single Molecule Fluorescent In Situ Hybridization (smFISH) on Whole-Mount Planarians.- Whole-Mount In Situ Hybridization in Large Sexual Schmidtea mediterranea.- Preparing Planarian Cells for High-Content Fluorescence Microscopy Using RNA In Situ Hybridization and Immunocytochemistry.- An RNA/DNA-Based Flow Cytometry Approach for Isolating Slow-Cycling Stem Cells.- ACME Dissociation-Fixation, Flow Cytometry, and Cell Sorting of Freshwater Planarian Cells.- Papain-Based Dissociation of Schmidtea mediterranea Cells.- Live Immunostaining and Flow Cytometry of Schmidtea mediterranea Cells.- Live Imaging in Planarians: Immobilization and Real-Time Visualization of Reactive Oxygen Species.- A Planarian Model System to Study Host-Pathogen Interactions.- TUNEL Staining in Sections of Paraffin-Enabled Planarians.- Quantitative Analysis of Planarian Pigmentation.- mRNA Transfection of S. mediterranea Cells for Luminescence Analysis.
