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Formaldehyde‐based whole‐mount in situ hybridization method for planarians
Author(s) -
Pearson Bret J.,
Eisenhoffer George T.,
Gurley Kyle A.,
Rink Jochen C.,
Miller Diane E.,
Sánchez Alvarado Alejandro
Publication year - 2009
Publication title -
developmental dynamics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.634
H-Index - 141
eISSN - 1097-0177
pISSN - 1058-8388
DOI - 10.1002/dvdy.21849
Subject(s) - planarian , biology , in situ hybridization , computational biology , microbiology and biotechnology , in situ , regeneration (biology) , gene expression , genetics , gene , physics , meteorology
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. © 2009 Wiley‐Liss, Inc.