I've been trying to dissect mouse epiblast at 5.5 dpc, and am unable to even see and differentiate the embryo from the decidua. I repeatedly end up destroying the decidua. The biggest problem is, I don't really know what I have to look for, if the embryo looks any different than the rest of tissue around, different colour, any specific site to trace it, where exactly to start dissecting from?
I want to generate EpiStem cell lines from these embryos. Can you please also suggest if I should use only PBS as a dissecting media or PBS mixed with a bit of my culture media as well? Any video links would also serve very helpful. Thanks a lot!
Dear Saleh,
Thanks a lot for your references. I am using the second reference as my protocol. Its just that, I can't see the epiblast in the 5.5 dpc stage. I tried the 6.5 dpc and because of increased vascularization, I am able to detect and obtain the epiblasts; but not at 5.5dpc. So I was hoping to get any advice you could give from your own personal experience.
Many thanks.
Regards,
Pooja
Dear Yunlong,
Thanks for your reference as well. I am currently using the same book.
Dear Pooja,
I know it is not easy work to dissect E5.5 mouse embryos at the beginning. Considering your difficulty in finding the embryo within decidua, I would recommend that you use GFP (or other fluorescence available)-labelled embryos and try to recognize (find) the embryos using fluorescent signal as a preliminary practice.
Good luck in your research.
Dear Yeonsoon,
Thank you for your reply. I did consider doing it earlier, but instead started back tracking- like dissect 6.5-->6.0-->5.75-->5.5 dpc. I have to admit its not going as well, but if it doesn't work, I shall try the GFP staining method. Thanks again.
Regards,
Pooja
- Badges
- Science topic
- Similar topics
- Medicine
- Disease
- Disease Models
- Animal Models
- Rodent Models
- Mouse Models