3/14/2017: Week 4 – Amnion, Amnion, Amnion
Hey, everyone!
I've got a few different things to talk about for my Week 4 post, and I'll get right into it.
BARNWATCH AND CHEYENNE
As I said in my last post, I had to spend the night at the ranch two Fridays ago watching Cheyenne and Lansdowne (mainly Cheyenne). Staying up late into the night was quite an interesting experience. Although my shift was only until 2 A.M., I ended up watching the horses until 4 A.M. with Nikash, whose shift was from 1 A.M. to 4 A.M. We had a pretty nice setup, with my laptop playing YouTube videos sitting right next to the laptop with live camera footage of the horses, and didn't observe anything unusual. But that was only until 3 A.M., when Cheyenne suddenly got down on the ground and started shaking and rolling quite violently. Because the two signs that horses are about to go into labor are either their water breaking or them lying down, we thought it was about to happen and ran to wake our mentor up. However, it ended up being nothing more than a very intense false alarm.
Cheyenne did give birth the next night! Unfortunately, I was out of town and therefore wasn't there to see it happen. However, I do have a picture.
Collection from Cheyenne went really well, and I'll talk more about what we did with that material in my Week 5 post. Lansdowne is still on yellow alert, and the team is keeping their fingers crossed that she'll give birth soon!
LIFE ON THE RANCH
Afte watching the horses until 4 A.M., Nikash and I headed off to a spare bedroom Mr. Ames had and got a few hours of sleep before ranch duty started. After waking up to a breakfast of cheese omelets and banana pancakes, we were back to work!
Surprisingly, ranch duty didn't involve anything new in Week 4. All our time was spent spraying for weeds around the whole ranch (since it had been almost a month since the last spraying) and cleaning both Lansdowne and Cheyenne. Strangely (and somewhat annoyingly), even though Cheyenne was much more sensitive at the time (being much closer to giving birth), she didn't give us any trouble at all, but Lansdowne was very uncooperative while we were trying to wash her. On a side-note, I managed to rip a hole in my jacket that day.
BACK TO THE LAB ONCE AGAIN
Now, for the most exciting part of this post and the reason behind the title: working with amnion.
Now that we had finished the first main step concerning the amniotic fluid, it was time to work with the amniotic membrane. Let me tell you, the membrane is some weird stuff. Because the baby horses are so large, there's a lot of material and it's very stretchy. Here's a picture so you can see it for yourself:
Keep in mind that the amnion is not in little pieces. It looks like a collection of blobs in the picture but it would look more like a huge sheet if everything in the bucket was all stretched out at once.
The first step of processing the amnion involved cutting it into small pieces (about 12" x 10") and scrubbing them with a sponge to get any dirt off. The amnion was also previously stored in a saline solution containing antibiotic to kill any bacteria.
Due to confidentiality issues, I can't talk about the next few steps in detail, but it essentially involved stretching out the small pieces of amnion that we cut out and drying them.
After we had the dry amnion ready, we stored them in some flat airtight bags and put some weight on them to keep them as flat as possible. That's pretty much where we've gotten with the amnion so far, and have repeated these steps many times trying to get through the tons of amnion we have to work with.
In my Week 3 post, I mentioned that we left some plates in the incubator to check for bacterial contamination. The control plate that we left open in the hood while working with the cells was completely bacteria free, telling us our working environment was sterile. This plate has a bit of a longer story behind it:
In case you don't remember what the four quadrants were indicative of, here's a reminder:
1) Antibiotic-treated cell suspension solution
2) Unfiltered amniotic fluid
3) Once filtered amniotic fluid
4) Twice filtered amniotic fluid
Bad news: as you can see in the picture, the cells were still very dirty, containing many different types of bacteria even after being treated with antibiotics. The good news is that the filters worked very well. Clearly, the unfiltered quadrant has a lot of bacterial growth, but as you can see, both filtered quadrants have no growth at all, indicating the amniotic fluid is now completely clean. Exciting stuff!
ADDITIONAL NOTES
I realize I'm a bit behind with my posts, and I'm sorry for that, but my Week 5 post will be out soon, and hopefully, everything will sync back on track after that. Week 5 was extremely busy, so the next post will likely be a long one. Thanks for reading, and keep your eyes open for the next post! One step closer to manufacturing the final products!
I've got a few different things to talk about for my Week 4 post, and I'll get right into it.
BARNWATCH AND CHEYENNE
As I said in my last post, I had to spend the night at the ranch two Fridays ago watching Cheyenne and Lansdowne (mainly Cheyenne). Staying up late into the night was quite an interesting experience. Although my shift was only until 2 A.M., I ended up watching the horses until 4 A.M. with Nikash, whose shift was from 1 A.M. to 4 A.M. We had a pretty nice setup, with my laptop playing YouTube videos sitting right next to the laptop with live camera footage of the horses, and didn't observe anything unusual. But that was only until 3 A.M., when Cheyenne suddenly got down on the ground and started shaking and rolling quite violently. Because the two signs that horses are about to go into labor are either their water breaking or them lying down, we thought it was about to happen and ran to wake our mentor up. However, it ended up being nothing more than a very intense false alarm.
Cheyenne did give birth the next night! Unfortunately, I was out of town and therefore wasn't there to see it happen. However, I do have a picture.
 |
| Cheyenne and Her Colt |
Collection from Cheyenne went really well, and I'll talk more about what we did with that material in my Week 5 post. Lansdowne is still on yellow alert, and the team is keeping their fingers crossed that she'll give birth soon!
LIFE ON THE RANCH
Afte watching the horses until 4 A.M., Nikash and I headed off to a spare bedroom Mr. Ames had and got a few hours of sleep before ranch duty started. After waking up to a breakfast of cheese omelets and banana pancakes, we were back to work!
Surprisingly, ranch duty didn't involve anything new in Week 4. All our time was spent spraying for weeds around the whole ranch (since it had been almost a month since the last spraying) and cleaning both Lansdowne and Cheyenne. Strangely (and somewhat annoyingly), even though Cheyenne was much more sensitive at the time (being much closer to giving birth), she didn't give us any trouble at all, but Lansdowne was very uncooperative while we were trying to wash her. On a side-note, I managed to rip a hole in my jacket that day.
BACK TO THE LAB ONCE AGAIN
Now, for the most exciting part of this post and the reason behind the title: working with amnion.
Now that we had finished the first main step concerning the amniotic fluid, it was time to work with the amniotic membrane. Let me tell you, the membrane is some weird stuff. Because the baby horses are so large, there's a lot of material and it's very stretchy. Here's a picture so you can see it for yourself:
 |
| Bucket of Amnion |
Keep in mind that the amnion is not in little pieces. It looks like a collection of blobs in the picture but it would look more like a huge sheet if everything in the bucket was all stretched out at once.
The first step of processing the amnion involved cutting it into small pieces (about 12" x 10") and scrubbing them with a sponge to get any dirt off. The amnion was also previously stored in a saline solution containing antibiotic to kill any bacteria.
 |
| Small Cut Portion of Amnion |
 |
| Wax On, Wax Off |
 |
| Stretched Amnion |
 |
| Dry Amnion |
After we had the dry amnion ready, we stored them in some flat airtight bags and put some weight on them to keep them as flat as possible. That's pretty much where we've gotten with the amnion so far, and have repeated these steps many times trying to get through the tons of amnion we have to work with.
In my Week 3 post, I mentioned that we left some plates in the incubator to check for bacterial contamination. The control plate that we left open in the hood while working with the cells was completely bacteria free, telling us our working environment was sterile. This plate has a bit of a longer story behind it:
 |
| Complicated? Plate |
In case you don't remember what the four quadrants were indicative of, here's a reminder:
1) Antibiotic-treated cell suspension solution
2) Unfiltered amniotic fluid
3) Once filtered amniotic fluid
4) Twice filtered amniotic fluid
Bad news: as you can see in the picture, the cells were still very dirty, containing many different types of bacteria even after being treated with antibiotics. The good news is that the filters worked very well. Clearly, the unfiltered quadrant has a lot of bacterial growth, but as you can see, both filtered quadrants have no growth at all, indicating the amniotic fluid is now completely clean. Exciting stuff!
ADDITIONAL NOTES
I realize I'm a bit behind with my posts, and I'm sorry for that, but my Week 5 post will be out soon, and hopefully, everything will sync back on track after that. Week 5 was extremely busy, so the next post will likely be a long one. Thanks for reading, and keep your eyes open for the next post! One step closer to manufacturing the final products!
The ranch stories are so exciting!! Glad you are enjoying your SRP!
ReplyDeleteThank you for the support!
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