Chicken embryo model? - (Apr/24/2007 )
I have a friend who is dealing with chicken embryo, and he is having some problem with incubating the chicken embryo... can someone help here?
He is actually doing an experiment which requires him to incubate fertilized egg. he needs to culture a fertelized egg in vitro to look for blood vessels.. and he is having problem in getting blood vessels...:
1. Break fertilized eggs, making sure that the egg yolk is not broken, I put the fertilized egg into a petri dish
2. Incubate in 36 degree celcius and observe the blood vessels growth
However, no blood vessels was seen, and the worse thing is the yolks ruptured by day 4-5 of incubation...
I was wondering how should incubate the eggs? Do I need to incubate the eggs for 3 days prior to breaking the shell? How long does it take for the blood vessels to develop? Thanks.
Hi Sanjuin,
The times I've seen this sort of procedure done, the eggshell was cracked at the top just enough to pull away a small part of the shell and make a window about a centimeter in diameter. The window was then covered with transparent saran film (Saran wrap). The idea was to minimize bacterial contamination of the embryo. I would think that the Petri-dish protocol you describe would inoculate the embryo with a lot of bacteria. You might try this less invasive approach. You can see a surprising amount of embryonic detail through the tiny window by illimunating the embryo through the eggshell with a bright lamp (a halogen fiberoptic light is great if you have it available).
Best of luck,
- Jon
Jon is right, there is no need to break the egg into a petri dish.
I used to work with chicken embryo (from E2 until E6) and the embryos were mostly fine. I would start to observe from E2 - E3 on. For more information I would check "Development of the chick embryo".
About the breaking thing, this is how I did it (Lets assume the embryo is 2 days old):
1. Put the fertilized egg on the side (the boxes the eggs are delivered in are pretty usefull as a pedestal).
2. Use a smal forceps to poke a tiny hole on the upper side of the egg. Carefully break away small parts of the chell until you have a window big enough to observe the embryo. right now it should look like a matchstick. You should see some somites and the "head" with the eye vesicles.
3. Use a syringe to remove about two to four ml of egg white. Some people make a small hole one the blunt side of the egg to do that.
4. Smear the edges of your window with silicone paste and close the window with saran wrap. This way it should be pretty tight.
5. Put the egg (on the pedestal) back into the incubator.
If you want I can send you a pic how it should look like.
Good luck!
He is actually doing an experiment which requires him to incubate fertilized egg. he needs to culture a fertelized egg in vitro to look for blood vessels.. and he is having problem in getting blood vessels...:
1. Break fertilized eggs, making sure that the egg yolk is not broken, I put the fertilized egg into a petri dish
2. Incubate in 36 degree celcius and observe the blood vessels growth
However, no blood vessels was seen, and the worse thing is the yolks ruptured by day 4-5 of incubation...
I was wondering how should incubate the eggs? Do I need to incubate the eggs for 3 days prior to breaking the shell? How long does it take for the blood vessels to develop? Thanks.
I used to work with chicken embryo (from E2 until E6) and the embryos were mostly fine. I would start to observe from E2 - E3 on. For more information I would check "Development of the chick embryo".
About the breaking thing, this is how I did it (Lets assume the embryo is 2 days old):
1. Put the fertilized egg on the side (the boxes the eggs are delivered in are pretty usefull as a pedestal).
2. Use a smal forceps to poke a tiny hole on the upper side of the egg. Carefully break away small parts of the chell until you have a window big enough to observe the embryo. right now it should look like a matchstick. You should see some somites and the "head" with the eye vesicles.
3. Use a syringe to remove about two to four ml of egg white. Some people make a small hole one the blunt side of the egg to do that.
4. Smear the edges of your window with silicone paste and close the window with saran wrap. This way it should be pretty tight.
5. Put the egg (on the pedestal) back into the incubator.
If you want I can send you a pic how it should look like.
Good luck!
Actually sanjiun81 was asking for me. I'm the friend she mentioned in the post... well, currently I'm still using the breaking egg method to cultivate the blood vessels... ur method sounds very interesting and less invasive, wanna ask, how long do i need to incubate before breaking a small window? I tried incubating at 37 degrees for 72 hrs ( or probably less coz I incubated on friday evening and broke them on monday morning) but only some developed embryo with the blood vessels... I need to put a disk filled with treatment to the vessels bed to see if there's any inhibition to the formation of blood vessels but so far, I've failed to get any results.
The window u mentioned, poke where? the pointed apex or the blunt end of the egg? for ur info, the eegs I purchased came seated on the egg tray with the blunt end faces the top, so when i incubate, i incubate with the blunt end facing upward... but to make window? which end of the egg? WHen u break, do u see a layer of white membrane covering the yolk? how do u get rid of the membrane? just peel it? I would be glad if u show me the photo u mentioned earlier. Mail to wengyw@yahoo.com, thanks alot... what's E2 and E6 by the way?? cheers...
Errr...thats a lot of questions 
First of all: Did nobody introduce you to the chick model? Because many of your questions should have been answered easily by your advisor.
The fertilized eggs are normally stored in a special fridge (I think it was 11 °C) until you use them. To incubate them we normally put them on the side in a humid chamber with a special mechanism to constantly roll the eggs (though some old incubators don't do this).
E2 means embryonic day 2, meaning the fertilized egg has been incubated for 48 hours (I never saw blood vessels at that point). For a more precise schedule of the development check the famous Hamburger/Hamilton-diagram.
Since I put my eggs on the side, the window was poked into the upper side of the egg. The thin white membrane was removed using a very thin glass pipette, I usually pulled myself.
PM me your email adress and I will send you some pics. Besides you really should sit together with your advisor, because it just doesn't seem normal to me that you are supposed to work with chick without having been introduced to any common knowledge about it.
Cya
Haplo
The window u mentioned, poke where? the pointed apex or the blunt end of the egg? for ur info, the eegs I purchased came seated on the egg tray with the blunt end faces the top, so when i incubate, i incubate with the blunt end facing upward... but to make window? which end of the egg? WHen u break, do u see a layer of white membrane covering the yolk? how do u get rid of the membrane? just peel it? I would be glad if u show me the photo u mentioned earlier. Mail to wengyw@yahoo.com, thanks alot... what's E2 and E6 by the way?? cheers...
I used to work with chicken embryo (from E2 until E6) and the embryos were mostly fine. I would start to observe from E2 - E3 on. For more information I would check "Development of the chick embryo".
About the breaking thing, this is how I did it (Lets assume the embryo is 2 days old):
1. Put the fertilized egg on the side (the boxes the eggs are delivered in are pretty usefull as a pedestal).
2. Use a smal forceps to poke a tiny hole on the upper side of the egg. Carefully break away small parts of the chell until you have a window big enough to observe the embryo. right now it should look like a matchstick. You should see some somites and the "head" with the eye vesicles.
3. Use a syringe to remove about two to four ml of egg white. Some people make a small hole one the blunt side of the egg to do that.
4. Smear the edges of your window with silicone paste and close the window with saran wrap. This way it should be pretty tight.
5. Put the egg (on the pedestal) back into the incubator.
If you want I can send you a pic how it should look like.
Good luck!
Actually sanjiun81 was asking for me. I'm the friend she mentioned in the post... well, currently I'm still using the breaking egg method to cultivate the blood vessels... ur method sounds very interesting and less invasive, wanna ask, how long do i need to incubate before breaking a small window? I tried incubating at 37 degrees for 72 hrs ( or probably less coz I incubated on friday evening and broke them on monday morning) but only some developed embryo with the blood vessels... I need to put a disk filled with treatment to the vessels bed to see if there's any inhibition to the formation of blood vessels but so far, I've failed to get any results.
The window u mentioned, poke where? the pointed apex or the blunt end of the egg? for ur info, the eegs I purchased came seated on the egg tray with the blunt end faces the top, so when i incubate, i incubate with the blunt end facing upward... but to make window? which end of the egg? WHen u break, do u see a layer of white membrane covering the yolk? how do u get rid of the membrane? just peel it? I would be glad if u show me the photo u mentioned earlier. Mail to wengyw@yahoo.com, thanks alot... what's E2 and E6 by the way?? cheers...
Wow... nice to see there are some progress in this discussion!!!!!
Good luck in your experiment, weng yew!!!