Week 10

This was my last week. I only went on Tuesday. I spent about 2 hours taking pictures of my results. There is a microscope attached to a computer which we can take pictures from. I got some good results and I can't wait to show them in my presentation.

Week 9 and Protocols

This week I did a Fluorescent Immunohistochemistry. It's basically the same procedure except the secondary antibody is fluorescent, the primary antibody is a lot more concentrated, the tissues must be kept in the dark after the secondary is added, and a few other things. I made a mistake during the experiment. Generally, we can put two antibodies together but only if they are made in two different animals. I was targeting tau and amyloid proteins so I put both their antibodies in. After 2-3 hours I realized that both antibodies were made in mouse (so I wasn't supposed to put them in together because then the secondary could bind to either and it would not differentiate) so I washed the tissue as much as I could in hopes that the antibody hadn't blinded yet. Then I put only one antibody in for amyloid plaques. Hopefully the results are okay. I briefly looked at them under the microscope and it seemed fine.

Week 8

I cover slipped my tissue this week:
     Steps for cover slipping:

•      Dehydrate the slides by putting them in 2 solutions of 70% ethanol, 2 solutions of 95%      ethanol, and 2 solutions of 100% ethanol for 5 minutes each
•      Put the slides in 3 solutions of xylene for 5 minutes each (xylene is in the fume hood)
•      Take a slide out of the last xylene with forceps and wipe the back
•      Smear a line of DPX along the slide and carefully slip the cover on the slide
•      Use the back of the forceps to push bubbles out 

Do one slide at a time.

I also completed a successful Nissl stain, and will work on analyzing that next week.

Week 7

I started my experiment. Everything flowed nicely and it was a successful experiment. Unfortunately, the person who gave me the tissue accidentally gave me two control tissues from the hippocampus instead of one control and one severe AD which means that I can't compare the tangles and plaques in control and SAD in the hippocampus. I will instead do a Nissl staining which will show the different sections of the hippocampus. I might be able to get actual SAD tissue from the hippocampus (but mostly likely not) and then I'll do the immuno on it if I get it.

These are my results from the IHC I did this week.