After several months, I had accumulated enough pig arteries to start the purification process. This was a big deal and an important milestone for my work. It is kind of like having enough ingredients to bake a cake or make a soufflé. A good cook will get all the ingredients together and work hard to mix and make the soufflé. You need enough of all the ingredients before you get started, and I now had all I needed. I had hundreds of frozen arteries collected over many weeks and would be using them all at one time. Protein purification is also a lot like making a soufflé. The whole soufflé can be ruined suddenly and with protein purification that is just as possible. However, I couldn’t just go get more ingredients from a grocery store. If I made a mistake doing protein purification it would take weeks more of collection and work to try again. Plus the purification process itself would take many steps taken over 3 months of time. As with the soufflé, I would not know until the very end of the process if I had been successful or not.

For the blending step of protein purification, the lab blender was a much larger version of the blender I had in my kitchen, with a much stronger motor and a metal container with a lid that would screw on. The screw-on lid was important because you did not want the chemical contents of a lab blender spraying across the lab due to a lid popping off. The motor was so powerful that the frozen arteries could get hot very quickly from the friction of the blender’s blades. After blending the arteries, I poured the resulting liquid into a big glass beaker and it looked like and had the consistency of a vanilla milkshake.

All the next steps in the purification needed to be done in the cold and the university had a whole room cooled to the temperature of a refrigerator. It was aptly called “the cold room.” I set up a bunch of plastic and glass tubing with the milkshake flowing through it in there. The whole setup went from the floor to the ceiling, about 10 feet high. At the end of the tubing a trickle of product would be produced at an agonizingly slow rate. The milkshake would flow through this system for a week, drop by drop, to produce enough product for me to work on. One of the chemicals I used in this cold room had a real bad smell—like a combination of rotten eggs and vomit[1]. So I tried very hard to prevent that chemical from leaking and smelling up the cold room. The cold room was used by many different people and I needed to prevent a spill that might close the room or make people upset.

As I became more proficient with my research and experiments, I was able to do more and more simultaneous experiments to produce more and more data. I would regularly have protein purification going in the cold room, experiments in the NMR lab and function experiments in the physiology lab all at once. The collection of data became a full time job and an obsessive mission of mine. It gave me a kind of thrill to be out on a date with Ann and know that I had three different experiments on going at the same time. Ann and I were becoming very close after about 3 months of dating, and I was falling very much in love with her.

The multi and simultaneous experiments became a kind of house of cards for me and eventually it did indeed fall down. Very late one Wednesday evening I came into the cold room and immediately smelt a chemical spill. My milkshake flow-through system had started to leak, and the chemical was on the floor around my tubing. It had only been leaking for a couple of hours, so was not so serious as to require an emergency cleanup. But even from this small a spill, the smell was quite evident and it would take me hours to clean up.

The first thing I did was stop the leak, then I made sure the milkshake and other chemicals had not been contaminated, and then I set about the cleanup. Near the end of the cleanup (about 6:00 A.M.) Jack, a graduate student from a neighboring lab, came in. I explained to him what happened and that the smell was dissipating and should not affect his experiments. He was unconcerned and went about his experiments.

I was heading home from the cleanup as Dr. Dillon was coming in for the day. I told him what happened, what I did and who I told about it. He shrugged and said, “Those things are bound to happen. Are you heading home?”

“Yes, to shower and come back to take down the NMR experiment.”

Dr. Dillon nodded and headed to his office. I drove home, showered, dressed and grabbed my skateboard to return to the NMR lab. I often would skateboard to the lab during the workday because it was fun, good exercise, and actually a faster way to get around campus than by car.

At the NMR lab I told Ann about my night and day of catch-up on my experiment’s spill, and explained that I wouldn’t be able to see her today because I had a lot of work and sleep to catch up on. She nodded her approval, seeming distracted. I hoped she wasn’t mad at me. Something seemed to be bothering her. Sleep deprived and frazzled as I was, I was awake enough to know something was wrong, and it gnawed at me.

[1] The chemical in question is beta mercaptoethanol.