3D Immunofluorescent Image Colocalization Quantification in Mouse Epiblast Stem Cells

Joshua G Dierolf; Andrew J Watson; Dean H Betts

doi:10.1007/978-1-0716-2281-0_7

3D Immunofluorescent Image Colocalization Quantification in Mouse Epiblast Stem Cells

Methods Mol Biol. 2022:2490:69-79. doi: 10.1007/978-1-0716-2281-0_7.

Authors

Joshua G Dierolf¹, Andrew J Watson^#^{1

2

3}, Dean H Betts^#^{4

5

6}

Affiliations

¹ Department of Physiology and Pharmacology, Schulich School of Medicine & Dentistry, The University of Western Ontario, London, ON, Canada.
² Department of Obstetrics and Gynecology, Schulich School of Medicine & Dentistry, The University of Western Ontario, London, ON, Canada.
³ The Children's Health Research Institute (CHRI), Lawson Health Research Institute, London, ON, Canada.
⁴ Department of Physiology and Pharmacology, Schulich School of Medicine & Dentistry, The University of Western Ontario, London, ON, Canada. dean.betts@schulich.uwo.ca.
⁵ Department of Obstetrics and Gynecology, Schulich School of Medicine & Dentistry, The University of Western Ontario, London, ON, Canada. dean.betts@schulich.uwo.ca.
⁶ The Children's Health Research Institute (CHRI), Lawson Health Research Institute, London, ON, Canada. dean.betts@schulich.uwo.ca.

^# Contributed equally.

PMID: 35486240
DOI: 10.1007/978-1-0716-2281-0_7

Abstract

This chapter details 3D morphological topography of colony architecture optimization and nuclear protein localization by co-immunofluorescent confocal microscopy analysis. Colocalization assessment of nuclear and cytoplasmic cell regions is detailed to demonstrate nuclear and cytoplasmic localization in mEpiSCs by confocal microscopy and orthogonal colocalization assessment. Protein colocalization within mESCs, mEpiLCs, and mEpiSCs can be efficiently completed using these optimized protocols.

Keywords: Colocalization; Confocal microscopy; Cytoplasmic; Immunofluorescence; Nuclear; Primed pluripotency; mEpiSC.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Nucleus
Coloring Agents
Cytoplasm
Germ Layers*
Mice
Microscopy, Confocal
Mouse Embryonic Stem Cells*

Substances

Coloring Agents

Grants and funding

CIHR/Canada