June 30th - Dr. Diehl's Lab

Quite a bit of work done....Need to check what I can and can't write here. Some stuff could still not be out and I am not supposed to mention those things. Top secret, if I tell you, I would need to ....... :-)

We started off the day with well blended bovine brains in buffer solution. Brains must be cleaned of protective tissue and as many blood vessels as possible (luckily Jonathan had done this already). The mixture is centrifuged and the supernatant (the liquid on top) is collected since it contains the microtubules and motor proteins. A solution is added supernatant to polymerize the microtubules (which will contain motor proteins) and placed in warm water to incubate. Afterwards, it was slowly pipetted onto a sucrose solution, due to difference in densities, the polymerized microtubules sink below the sucrose. Once set up, it is centrifuged and pellet of microtubules at bottom is collected and washed with original buffer solution along with some polymerizing agent and then incubated and centrifuged again and pellet is collected. Pellet is then suspended in buffer, ...... , and ATP solution. The ATP provides energy for the motor proteins to separate from microtubules. Solution is incubated at 37 degrees C, which is about body temperature, when motor proteins are more active. Solution is centrifuged and separated into nine, 100 microliter samples. Eight of the samples are frozen in liquid nitrogen and one is going through analysis. The sample went through some more procedures (secret stuff :-) ) and then analyzed using a certain type of microscope. (again there are some things I cannot mention, so please don't ask.....)