The long and winding road to Eureka

Today I have had a new, awesome, big experiment planned that would take up for 12 hours, but would still, theoretically, be totally worth it. I wanted to demonstrate that a specific subset of white blood cells (WBCs) would display increased expression of an immunosuppressive enzyme, upon a certain treatment, that could potentially explain the survival of - otherwise rejected - transplanted organs we are observing in mice. I wanted to make a 07:00 to 19:00 day and my alarm was set for 06:00. The drama started when my sleep was brutally disturbed by the smoke detector that went off at 03:00 A.M. Luckily, it was a false alarm and the batteries were merely empty. Nevertheless, it was hard for my adrenalin-filled-body to fall back asleep again. Perhaps it is not the smartest thing to perform a twelve hour during experiment on a basis of four hours of sleep. But I was so excited for this experiment, which would lift my research to the next level, that I was still having a very optimistic vibe. Plus, it took me three weeks to book this machine that was required for today's experiment. That's not something I just wanted to give up! So, I started my morning at 07:00 to collect the spleens of my treated mice, and continued isolating the WBCs out of these spleens. I was only interested in a particular subset of these WBCs called dendritic cells. In order for me to obtain these cells through this booked machine, I first needed to get rid of cells I wasn't interested in. This is called selective enrichment. I depleted all the B and T cells by a technique I was not yet familiar with. I find it that learning a new technique requires a lot of focus and energy. Therefore I am glad that this wonderful substance called coffee was invented. After my successfully performed selective enrichment, I started staining my cells for markers which are specifically expressed on the cells I was interested in, these dendritic cells. This awesome machine I booked three weeks in advance has the ability to recognize these stained cells and is friendly, more importantly, advanced enough, to select these cells and give it back to me in a tube. I started smiling when I had my hands on these tubes filled with my favourite cells, isolated from all the other white blood cells. I was walking back to my bench while protecting my precious tubes. Ten hours had past, two to go. The most tricky part was behind me. Now I needed to proceed with demonstrating that these treated cells produce more immunosuppressive enzyme compared with untreated cells. Cells are like little factories producing thousands of wonderful proteins. This production process is read from the DNA, and transcribed into RNA. I needed to isolate the RNA from my gorgeous cells living in my tubes. During this process of RNA isolation I made a mistake I could not afford at 19:00 o' clock. Instead of giving them a pleasant concentration of 1x PBS solution in which cells will happily flow for efficient replacement from tube to tube, I gave them by accident a ten times stronger concentration. I can tell you that cells are not happy with that. They weren't happy at all. As a matter of fact, they were dying in these conditions. After eleven hours of delicate procedures to obtain my precious cells, I accidentally intoxicated them!! At the end of the day, I only had tubes with selectively sorted dead cells.

Fuck.