小鼠上胚层干细胞中的3D免疫荧光图像共定位定量分析

3D Immunofluorescent Image Colocalization Quantification in Mouse Epiblast Stem Cells.

作者信息

Dierolf Joshua G, Watson Andrew J, Betts Dean H

机构信息

Department of Physiology and Pharmacology, Schulich School of Medicine & Dentistry, The University of Western Ontario, London, ON, Canada.

Department of Obstetrics and Gynecology, Schulich School of Medicine & Dentistry, The University of Western Ontario, London, ON, Canada.

出版信息

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

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

PMID:35486240

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.

摘要

本章详细介绍了通过共免疫荧光共聚焦显微镜分析进行集落结构优化和核蛋白定位的三维形态拓扑学。详细阐述了核区和胞质区的共定位评估，以通过共聚焦显微镜和正交共定位评估来证明小鼠胚胎干细胞样细胞（mEpiSCs）中的核定位和胞质定位。使用这些优化方案可以有效地完成小鼠胚胎干细胞（mESCs）、小鼠胚胎外胚层干细胞样细胞（mEpiLCs）和小鼠胚胎干细胞样细胞（mEpiSCs）内的蛋白质共定位。