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Methods Mol Biol. 2018;1774:353-366. doi: 10.1007/978-1-4939-7802-1_10.

Fixation, Processing, and Immunofluorescent Labeling of Whole Mount Planarians.

Author information

1
Functional and Chemical Genomics Research Program, Oklahoma Medical Research Foundation, Oklahoma, OK, USA. david-forsthoefel@omrf.org.
2
Department of Biology, San Diego State University, San Diego, CA, USA.
3
Department of Integrative Biology, Howard Hughes Medical Institute, Morgridge Institute for Research, University of Wisconsin-Madison, Madison, WI, USA.
4
Department of Biology, San Diego State University, San Diego, CA, USA. rzayas@mail.sdsu.edu.

Abstract

Efforts to elucidate mechanisms of regeneration in the planarian Schmidtea mediterranea have included the application of immunocytochemical methods to detect specific molecules and label cells and tissues in situ. Here we describe methods for immunofluorescent labeling of whole mount planarians. We outline protocols for fixation and steps for processing animals prior to immunolabeling, incorporating commonly utilized reagents for mucus removal, pigment bleaching, tissue permeabilization, and antigen retrieval. Because processing steps can mask or degrade antigens, we also recommend protocol parameters that can be tested simultaneously to optimize sample preparation for novel antibodies.

KEYWORDS:

Antibodies; Flatworms; Histology; Immunostaining

PMID:
29916163
DOI:
10.1007/978-1-4939-7802-1_10
[Indexed for MEDLINE]

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