Schmidtea mediterranea: Methods and Protocols

ISBN-13: 9781071632741 / Angielski

Luca Gentile

Opis

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.

Spis treści

1. Building Phylogenies from Transcriptomic Data

            Lisandra Benítez-Álvarez, Laia Leria, Daniel Dols-Serrate, and Marta Riutort

 

2. Isolation and Library Preparation of Planarian piRNAs

            Iana V. Kim, Tim Demtröder, and Claus-D. Kuhn

 

3. Genome-Wide Analysis of Planarian piRNAs

            Andreas Pittroff, Iana V. Kim, Tim Demtröder, and Claus-D. Kuhn

 

4. Combining Fluorescent In Situ Hybridization with Immunofluorescence and Lectin Staining in Planarians

            Daniel Font-Martín, Eudald Pascual-Carreras, and Emili Saló

 

5. Colorimetric Whole-Mount In Situ Hybridization in Planarians

            Susanna Fraguas, Mª Dolores Molina, and Francesc Cebrià

 

6. Single Molecule Fluorescent In Situ Hybridization (smFISH) on Whole-Mount Planarians

            Elke F. Roovers and Kerstin Bartscherer

 

7. Whole-Mount In Situ Hybridization in Large Sexual Schmidtea mediterranea

            Miquel Vila-Farré, Hanh Thi-Kim Vu, and Jochen C. Rink

 

8. Preparing Planarian Cells for High-Content Fluorescence Microscopy Using RNA In Situ Hybridization and Immunocytochemistry

            Markus A. Grohme, Olga Frank, and Jochen C. Rink

 

9. An RNA/DNA-Based Flow Cytometry Approach for Isolating Slow-Cycling Stem Cells

            Nicole Lindsay-Mosher, Alyssa M. Molinaro, and Bret J. Pearson

 

10. ACME Dissociation-Fixation, Flow Cytometry, and Cell Sorting of Freshwater Planarian Cells

            Helena García-Castro, Elena Emili, and Jordi Solana

 

11. Papain-Based Dissociation of Schmidtea mediterranea Cells

            Claudia Ortmeier and Luca Gentile

 

12. Live Immunostaining and Flow Cytometry of Schmidtea mediterranea Cells

            Claudia Ortmeier and Luca Gentile

 

13. Live Imaging in Planarians: Immobilization and Real-Time Visualization of Reactive Oxygen Species

            Vincent Jaenen, Karolien Bijnens, Martijn Heleven, Tom Artois, and Karen Smeets

 

14. A Planarian Model System to Study Host-Pathogen Interactions

            Eli Isael Maciel, Ashley Valle Arevalo, Clarissa J. Nobile, and Néstor J. Oviedo

 

15. TUNEL Staining in Sections of Paraffin-Enabled Planarians

            Maria Rosselló and Teresa Adell

 

16. Quantitative Analysis of Planarian Pigmentation

            Matthew Pittendreigh, Kaleigh Powers, Meenalosini Vimal Cruz, and Jason Pellettieri

 

17. mRNA Transfection of S. mediterranea Cells for Luminescence Analysis

            Uri Weill, Richard Nelson Hall, Leonard Drees, Bo Wang, and Jochen C. Rink