Getting a leg up with lab work
After a few unsuccessful attempts at doing immunohistochemistry (IHC) on very teeny meniscal sections, I got the staining methodology down! I will spare you with the details of my tedious work, but for anyone learning how to do IHC, let me tell ya, its a lot like western blotting, which I did a ton of last year.
Here’s a brief summary of what I have been doing lately…
These are the samples on slides that I work with. The rabbit menisci are really small, and only 6micrometers thick. I use a PAP pen to outline each section, so that I can conserve the amount of liquid I use throughout the process (antibodies are really expensive…)
I have to be careful not to suck up the sections when I remove the liquid from the slides. Although the sections should stay firmly attached to the slides, sometimes the sections loosely fall off because of the digestions with hyaluronidase, pronase, and proteinase K that I use.
I don’t really mind having to do so much IHC this time of year, because the -43F windchill outside is pretty unattractive at the moment. I do have to walk across campus to use the fluorescent microscope though… BRR!