Journal article
A Method for Sectioning and Immunohistochemical Analysis of Stem Cell-Derived 3-D Organoids
Current protocols in stem cell biology, Vol.37(1), pp.1C.19.1-1C.19.11
05/12/2016
DOI: 10.1002/cpsc.3
PMCID: PMC4900235
PMID: 27171793
Abstract
This unit describes a protocol for embedding, sectioning, and immunocytochemical analysis of pluripotent stem cell-derived 3-D organoids. Specifically, we describe a method to embed iPSC-derived retinal cups in low-melt agarose, acquire thick sections using a vibratome tissue slicer, and perform immunohistochemical analysis. This method includes an approach for antibody labeling that minimizes the amount of antibody needed for individual experiments and that utilizes large-volume washing to increase the signal-to-noise ratio, allowing for clean, high-resolution imaging of developing cell types. The universal methods described can be employed regardless of the type of pluripotent stem cell used and 3-D organoid generated. © 2016 by John Wiley & Sons, Inc.
Details
- Title: Subtitle
- A Method for Sectioning and Immunohistochemical Analysis of Stem Cell-Derived 3-D Organoids
- Creators
- Luke A Wiley - Stephen A. Wynn Institute for Vision Research, Department of Ophthalmology and Visual Sciences, Carver College of Medicine, University of Iowa, Iowa City, IowaDavid C Beebe - Departments of Ophthalmology & Visual Sciences and Cell Biology and Physiology, Washington University School of Medicine, Washington University, St. Louis, MissouriRobert F Mullins - Stephen A. Wynn Institute for Vision Research, Department of Ophthalmology and Visual Sciences, Carver College of Medicine, University of Iowa, Iowa City, IowaEdwin M Stone - Stephen A. Wynn Institute for Vision Research, Department of Ophthalmology and Visual Sciences, Carver College of Medicine, University of Iowa, Iowa City, IowaBudd A Tucker - Stephen A. Wynn Institute for Vision Research, Department of Ophthalmology and Visual Sciences, Carver College of Medicine, University of Iowa, Iowa City, Iowa
- Resource Type
- Journal article
- Publication Details
- Current protocols in stem cell biology, Vol.37(1), pp.1C.19.1-1C.19.11
- DOI
- 10.1002/cpsc.3
- PMID
- 27171793
- PMCID
- PMC4900235
- NLM abbreviation
- Curr Protoc Stem Cell Biol
- ISSN
- 1941-7322
- eISSN
- 1938-8969
- Publisher
- United States
- Grant note
- DP2 OD007483 / NIH HHS R01 EY024588 / NEI NIH HHS R01 EY024605 / NEI NIH HHS
- Language
- English
- Date published
- 05/12/2016
- Academic Unit
- Iowa Neuroscience Institute; Ophthalmology and Visual Sciences
- Record Identifier
- 9983980062002771
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