Now that my main job is microdissection, I spend a lot of time using different microscopes. Only in passing will I note that most microscopes in my lab come with notes on them to remind us all of the basics of using anything; “turn off after use”, etc. And yes, I do have a favorite scope – the Leica Wild 10 in the embryology room. It doesn’t slip out of focus, is located in a room where I can turn off all the lights and shut the door, has appropriate levels levels of magnification (because I need to dissect little tiny things!), and nice functioning adjustable lights
Although I love the Leica Wild, it may soon be replaced with a new winning scope. Last week I went down down down into the basement and visited the BioRad MultiPhoton microscope for the first time. This microscope is so cool that it exists in a private room jammed with what I can only describe as tons of complicated stuff and what seems to be one helluva computer. According to the word on the street, this baby cost a ton of money . On top of that, it requires a full time guy (appropriately bearded and outfitted in a seemingly endless series of tucked in flannel shirts) to teach people like me how to get images out of it. This guy is not a tech (Editing from the future: I soon figure out he does MUCH more than just teach people like me to not break expensive equipment). He’s the director of Imaging and Physiology who helps teach classes with titles like “Three-dimensional Cryo Electron Microscopy of Single Particles”. And he has a wife and a two year old child – obviously amazing.
But as much as I would like to figure out exactly how one becomes expert in operating such machines, and in the process of three-dimensional cryo electron microscopy of single particles, while still finding the time and inclination to procreate – it will be a while until I get to take my little pulsating blobs of living tissue down to make movies of their insides. Crawling before walking… and really, before I even think about crawling I have got to flip myself over onto my belly and turn on some primal brainstem and spinal cord pattern generators to see if they’re working.
In terms of the project, this means I have to get the live tissue dissected out and properly labeled to see how certain axons grow during the different stages of development. Today was devoted to getting that live tissue dissected out. This involves taking one of those gorgeous embryos, removing it from the egg, eviscerating, decapitating, and “blocking” it so that only the spinal column (with T4 carefully marked) remains. Then begins the rather painstaking task of removing the neural tube, with dorsal root ganglia and (hopefully) dorsal roots attached. Amazingly, after plenty of frustration, and the acquisition of my very own personal pair of very pointy number five forceps, I was able to do it three times. And at five pm, my difficult to please mentor said “That’s nice and clean -but maybe too clean, you’ve probably severed the dorsal roots”.
Ah, science. Frustration, trial and error, and very picky people who notice every little thing. But, in any case, I left the lab floating on a minute cloud of satisfying accomplishment. Ready settle into the evening with a review paper on live imaging, or at least to carry it with me from room to room with good intentions.