Book chapter
Combining Whole-Mount In Situ Hybridization with Neuronal Tracing and Immunohistochemistry
In Situ Hybridization Methods, pp.339-352
Neuromethods, Springer New York
01/15/2015
DOI: 10.1007/978-1-4939-2303-8_17
Abstract
Despite the presence of several markers to study the expression of genes (eGFP, LacZ), in situ hybridization remains one of the most powerful techniques to analyze gene expression. While this allows cellular identification of the expression of a single gene and, using fluorescent in situ hybridization two or occasionally more genes, it is often necessary to combine this technology with assays of neuronal projection/morphology, protein expression using antibody staining, and histology for cytological details. Since each task has certain levels of false negatives, combining them in a single preparation can compromise further correlative studies due to loss of fluorescence, loss of antigenic epitope, or loss of tissue morphology. We have designed a protocol that, when performed in sequence, will enable the researcher to combine several of these technologies in the same sample saving time and sparing expense. By combining neuronal tracing, whole-mount in situ hybridization, immunohistochemistry, and histology one can extract a maximal amount of data with limited loss in fidelity of each technique and optimal data superposition for a more complete understanding of phenotypes.
Details
- Title: Subtitle
- Combining Whole-Mount In Situ Hybridization with Neuronal Tracing and Immunohistochemistry
- Creators
- Jeremy S Duncan - Division of Otolaryngology and Department of Neurobiology and Anatomy, University of Utah School of Medicine, Salt Lake City, USAKaren L Elliott - The Department of Biology, College of Liberal Arts and Sciences, University of Iowa, Iowa City, USAJennifer Kersigo - The Department of Biology, College of Liberal Arts and Sciences, University of Iowa, Iowa City, USABrian Gray - Molecular Targeting Technologies Inc., West Chester, USABernd Fritzsch - The Department of Biology, College of Liberal Arts and Sciences, University of Iowa, Iowa City, USA
- Resource Type
- Book chapter
- Publication Details
- In Situ Hybridization Methods, pp.339-352
- Series
- Neuromethods
- DOI
- 10.1007/978-1-4939-2303-8_17
- eISSN
- 1940-6045
- ISSN
- 0893-2336
- Publisher
- Springer New York; New York, NY
- Language
- English
- Date published
- 01/15/2015
- Academic Unit
- Iowa Neuroscience Institute; Biology; Craniofacial Anomalies Research Center
- Record Identifier
- 9984070891502771
Metrics
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