Jul 14, 2025
Pathological analysis of mosue embryos
- Peng Xu1,2,
- Caroline J. Zeiss3,
- Pietro De Camilli1,2
- 1Departments of Neuroscience and of Cell Biology, Howard Hughes Medical Institute, Program in Cellular Neuroscience, Neurodegeneration and Repair, Yale University School of Medicine, New Haven, Connecticut 06510, USA;
- 2Aligning Science Across Parkinson's (ASAP) Collaborative Research Network, Chevy Chase, MD, 20815;
- 3Department of Comparative Medicine, Yale School of Medicine, New Haven, Connecticut 06510, USA
Protocol Citation: Peng Xu, Caroline J. Zeiss, Pietro De Camilli 2025. Pathological analysis of mosue embryos. protocols.io https://dx.doi.org/10.17504/protocols.io.kxygx4emdl8j/v1
Manuscript citation:
Defect in hematopoiesis and embryonic lethality at midgestation of Vps13a/Vps13c double knockout mice
Peng Xu, Rubia Isler Mancuso, Marianna Leonzino, Caroline J. Zeiss, Diane S. Krause, Pietro De Camilli
bioRxiv 2025.05.09.653147; doi: https://doi.org/10.1101/2025.05.09.653147
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: July 12, 2025
Last Modified: July 14, 2025
Protocol Integer ID: 222380
Keywords: pathological analysis of mouse embryo, mouse embryo, embryo, pathological analysis
Funders Acknowledgements:
National Institutes of Health
Grant ID: NS36251
National Institutes of Health
Grant ID: DA018343
Aligning Science Across Parkinson
Grant ID: ASAP-000580
Abstract
A protocol for the pathological analysis of mouse embryos.
Troubleshooting
Pregnant dams were euthanized at estimated embryonic day 12.5 using a carbon monoxide chamber (filled gradually to 70%) followed by creation of pneumothorax.
The uterus was extracted in its entirety, after which fetuses (n=6 per litter) and their membranes were dissected intact from the uterus. The tail of each fetus was harvested for genotyping.
Fetuses were fixed in Bouin’s solution for 48 hours, followed by bisection along the sagittal plane. Cassetted tissues were submitted for standard paraffin embedding, processing, and generation of 20-30 5µm sections at 20µm intervals.
Sections were stained with hematoxylin and eosin.
Immunoperoxidase stains were performed using unstained 5µm paraffin sections using primary antibodies at a 1:100 dilution.
After deparaffinization and rehydration, antigen retrieval was performed using sodium citrate buffer (10mM Sodium Citrate, 0.05% Tween 20, pH 6.0) at 95-100oC for 10 minutes.
Immunostaining was performed using a Dako autostainer.
Label was visualized by 0.05% 3′,3′-diaminobenzidine (DAB) as a chromogen, precipitated by 0.01%
hydrogen peroxide.
Light microscopic images were taken using a Zeiss Axioskop and with Axiocam MrC camera.