Winter quarter is deep underway, and as a result sickness has permeated through the student body of UCSB. Unfortunately, my cloak of vitamin C, airborne, and continuous hand washing did not prove to have the power of invisibility, and sickness found me. All through my horror week of 4 midterms (OChem, Physics, Spanish 100, and Spanish 25) my voice had abandoned me, my throat ached from coughing, my body shivered with a 101 degree fever, and my only consolations were boxes of nyquil and sudafed. After two weeks without symptom relief, I finally convinced myself to take a trip to the urgent care doctor who gave me antibiotics that have finally kicked in, and kicked out whatever sinus infection had decided to keep my ears plugged for so long.
Today is Monday, and this is the first week I have felt human again. So I dropped into my research lab to check on the Celastrol brain stainings that I had somehow finished (in an incoherent state, no doubt) over the course of the past two weeks. Now, keep in mind that these stainings are supposed to begin with a primary antibody 48-52 hour incubation, continue with a secondary antibody 24-48 hour incubation, and conclude with an afternoon of tissue washing and the process of mounting the brain slices on slides. This process generally occurs, Friday, Monday, and then Wednesday mounting. So less than one week.
My Celastrol stainings have been sitting in incubation buffer (contains a soap to perforate the tissues and allow antibodies to enter individual cells) and antibodies for TWO weeks, not less than one. You might think: no big deal, the brains were sitting in soap, so they will just be extra clean! But, no. Soap + brain tissue = thousands of tiny holes in all the membranes and extremely delicate slices. I tried to mount them on the slides and most of them fell apart simply in the transportation process. It was an epic fail, and although some of the tissues remained intact, the experiment will have to be repeated. So that’s what I will be doing this Friday after the next Ochem midterm.
On the bright side, the tissues that survived the mounting process will probably appear beautifully stained. Due to the abundance of tiny holes in the membranes, the antibody should have had free access to the proteins inside the cells. I guess we will see.
Well, back to studying for my Spanish Linguistics midterm tomorrow!