No, I’m not talking about science classes (though those can sometimes be hard too). I’m talking about IRB-forms-signed-in-blood, strap-on-your-goggles-and-get-ready-to-kick-ass, thrown-out-to-sea-sucking-down-methanol laboratory science (and often its friendly cousin let’s-sit-behind-a-computer-and-spit-out-all-our-thoughts-into-SPSS dry lab science, but I’m quickly discovering that these are two games that use completely different racquets).
So you’ve finally gotten you hands on your very own little pilot study, with a promising substrate that hints at the central nervous activity about which you hope to spend a career learning. You’ve tabulated the literature, developed your hypothesis, formulated your methods, secured your materials, cleared your plans with your P.I. You’re pumped. You’re psyched. Your palms are sweating, not least of all because they’re crammed into latex gloves (which, along with your lab coat and goggles, are still super exciting to put on every time).
Your very first step at the bench is child’s play. Dissolve your substrate in a solvent at a concentration that won’t kill your mice upon injection. You brush off the dusty high-school-science data bank sitting in the back of your brain and get calculating off the lab standard for injection volume and your study’s desired doses. The math is tedious but straightforward. Your gloves, coat, and goggles, which you’ve been wearing everywhere for the last week,* protect you from wayward paper and splashing ink.
Finally armed with a list of desired solute masses, you proceed to dissolve the ingredients as directed in the most authoritative paper you have. You measure out your powder by the tenth of a milligram, painstakingly pipet the right amount of solvents by the microliter. An artist who has just produced her first masterpiece, you step back to admire your work… which isn’t dissolving.
Okay, no sweat. Reagent conditions can vary by lab. The literature suggests at least two more options. You’ll try those before getting too picky about temperature and pH buffers. Half an hour later, you step back to admire your three masterpieces…
…none of which is dissolving.
You dive once more into literally every single paper on PubMed that mentions the little f*cker you need so desperately to dissolve. Of course, only like three out of the hundred were performed under conditions relevant to your experiment.
Congratulations! You have left the recipe kitchen and entered the world of science.
And it’s hard.
You can’t use just any solvent; the organic reagent precipitates at a sneeze. You can’t just pump it into our friends DMSO or ethanol; mice can handle only very low concentrations of organic solvent. You can’t win the game by deluging a little bit of solute with a ton of solvent because you shouldn’t be injecting more than 300 microliters of solution into your mouse at once and you want your administered solute masses to have logarithmic separation over a large range. And you can’t trick the reagent into dissolving by increasing the solution temperature too much because you don’t want to incite any mystery reactions.
But you are confident that optimism and logic will build something great within even these tall and labyrinthine walls. You narrow your goggled eyes, button up your lab coat, snap your gloves farther up your wrist (just for the hell of it), and set to work calculating, massing, adjusting, heating, pipetting, mixing, and creating, this time to the tune not of a published protocol but the rational symphony of your own neurons firing over accumulated experiential data.
And, finally, standing over your finished product nearly twenty-four hours after you first anticipated problem number one would be solved, you’re left wondering why you never had so much trouble in any of your gazillion pre-med lab science courses. Oh, yeah. Because the TF did all the hard stuff.