Another day another dollar, a new week a new protocol. I got to explore a revised protocol for harvesting lymphocytes from blood, which was as much fun as it was frustrating. I also got to learn a little about Rapamycin and its antibiotic properties – it happens to change the color of muscle from the typical fleshy red to a still very natural goldfish orange when applied to open wounds. I recently found a good centrifuge companion to fill the time when I actually have 30 minutes to spare during my busy day; Donna Tartt’s The Goldfinch has been a good read so far.
(Figure 1. Hail Mary full of grace… A hand homogenizer from Braun, Germany for your viewing pleasure. There really isn’t much to it.)
Executing a new protocol leaves me with very little to say, but I’m certain now that cell harvesting/culturing is surely the bane of biologists and researchers everywhere. That and grant submissions. What I have done so far is draw more blood and separate the lymphocytes once again, but this time I’m going through the tedious process of hand homogenizing the cells in 60 delicate strokes. One good stroke takes about 40 seconds, just to give you a sense of the leisurely process. Storage is also quite different for this new sample set. I have had to make up several new buffers, the most exciting one containing a trace amount of DMSO – used to keep protease inhibitors in solution. This is an odd catch-22 though. The protease inhibitors that inhibit the enzymes in the cells from proteolysis (this hydrolyzes the peptide bonds keeping proteins intact, which is important since anyone studying biology knows protein function directly correlates to protein structure, i.e. peptide bonds) are made in solution with DMSO which can potentially denature said proteins in cell samples. Get what I mean?
Despite all of the new obstacles, I am happy to tell you – dearest reader – that the process has been completed and the cell samples are sitting in -80°C, respectfully adhering to the protocol. The last part, called for flash freezing the cells in a bath of liquid nitrogen. I thought it wouldn’t be a huge deal, after all Reed has it’s own Nitrogen Day Czar’s and liquid nitrogen ice cream tradition, it’s not like I’m inexperienced with the compound.
(Figure 2. A coworker got a set of some research jargon magnets at a seminar. This struck me the most. Poetry to come.)
What I have learned or been reminded of so far:
1. DMSO can make your skin itch and may also leaves an unusual savory flavor in your mouth after you come in to contact with it (Chem202 students can put that on their list with isoamyl acetate).
2. Cells are very uncooperative when sustained outside your body; I am repeatedly amazed that I’m able to work with them at all.
3. Mice travel very slowly…