Journal
DEVELOPMENTAL DYNAMICS
Volume 238, Issue 2, Pages 443-450Publisher
WILEY
DOI: 10.1002/dvdy.21849
Keywords
in situ; whole-mount; planaria; mucolytics
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Funding
- Damon Runyon Foundation
- NIH Developmental Biology Training Grant
- NIH-NIGMS
- European Molecular Biology Organization
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Whole-mount in situ hybridization (WISH) is a powerful tool for visualizing gene expression patterns in specific cell and tissue types. Each model organism presents its own unique set of challenges for achieving robust and reproducible staining with cellular resolution. Here, we describe a formaldehyde-based WISH method for the freshwater planarian Schmidtea mediterranea developed by systematically comparing and optimizing techniques for fixation, permeabilization, hybridization, and postprocessing. The new method gives robust, high-resolution labeling in fine anatomical detail, allows co-labeling with fluorescent probes, and is sufficiently sensitive to resolve the expression pattern of a microRNA in planarians. Our WISH methodology not only provides significant advancements over current protocols that make it a valuable asset for the planarian community, but should also find wide applicability in WISH methods used in other systems. Developmental Dynamics 238:443-450, 2009. (c) 2009 Wiley-Liss, Inc.
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