Axons Week 1: Fungus is Among Us

Happy June everyone!

Last week, we started by growing up some HEK293 cells. They’re a cell line we’re using because they’re really good at growing proteins. We want to eventually use these cells to package a virus (don’t worry, the virus is replication incompetent). We will then use the virus to “edit” the genome of neurons to get rid of the SARM protein (see my first post for the abstract if you’re totally lost right now, or even email me!).

Before we do that though, we need to figure out what density of those cells most efficiently makes virus. Our goal for the first week was to carry out a couple of experiments to test cell density and transfection DNA concentration.

However, we ran into a bit of a snag. Pro-tip: your incubator water bath should never look like this:

Gross.

Gross.

We had a fungal contamination. And it was terrible. And we had to throw out most of our cells.

We suspected it was from the micropipettors, which can sometimes harbor fungus or bacteria in their shafts. So we gathered all of them up along with the shelves and pan from the incubator and autoclaved everything (an autoclave chamber reaches super high temperature and pressure, so we use it to disinfect things).

Thankfully, we still got some usable data on cell density and transfection efficiency. Now we can carry on this week and get really good at transfections before we try to make the actual virus.

This coming week, I’m hoping to get some good retinal ganglion explants, too. Those neurons are what we’ll use when we actually get the virus going. I’ll also be getting certified to use the confocal microscope, which is huge and uses lasers to get really detailed images.

Other than that, I’ve been helping take care of the zebra finches and enjoying the relaxed atmosphere on campus. It’s amazing to not feel stress emanating from all sides for once. And Swem? Empty. It’s a beautiful time to be here, y’all.