In science, sometimes the only way we can actually reach our destination is with all of the hidden failures along the way that guide us to the end game. I guarantee every scientist will experience their fair amount of failure, but all of it will make you better. In optimization, essentially the entirety of my job, this is especially prevalent. When figuring out the best procedure, you have to go through a lot of failed ones. In addition to the one successful procedure and the one in progress, we now have yet another to optimize. Last week, we started a procedure to extract bacterial DNA from human stool samples. We ran a gel to see if the DNA extraction was successful. Our extraction procedure was utterly unsuccessful. Though it would have been nice to find success on the first try (something I now realize was wishful thinking) it is going to make the end result much more of a success. Without stumbles along the way, we can never get better.
In continuation with our overall project, we have been continuing to prepare slides to look at under the microscope. If we find that the SERT levels are not the reason behind high levels of 5-HTT, we will have to begin optimizing a second IHC procedure for a different protein. I am very excited to optimize the second procedure. So far, we have not decisively concluded our results. We still need to learn how to use the confocal microscope so we can see both contrasts at the same time and exposure. I expect we will begin using the confocal soon. As for the slides, our results have gotten even better. In up-ing the exposure time for 20ms to 40ms, we were able to see the SERT much more clearly and out edits came out much better. I have attached a picture above. One interesting thing we noticed on our last round of slides was the distal colon had much higher levels of SERT than any other portion of the intestine. This was very strange because we usually have the highest levels in the ileum. We will continue to examine this in further slides.
Over the past two weeks, the lab members have truly begun to become my friends. This experience has been incredible and although I am sad to only have two weeks left, I am really looking forward to finishing my project here! Also this week, Ms. Cozine came to Washington, D.C. and had lunch with me and my grad student. I was able to show her the microscope we currently use, which I know get to operate myself. We had a great time and I really appreciate her coming!
Good luck with your next two weeks! If you have time, read Michelle's blog - she is coming to the same conclusion that you are - it takes lots of trial and error to optimize your results. Hope your tennis matches go well!
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