Student Research at Duke

This past week, I have learned that it is important to know what chemicals you are dealing with- their toxicity, the precautions suggested, and the method of disposal. Dr. Calakos emphasized that one of the first things to do when working with a new substance is to look up its MSDS on the Duke OESO website. Two of the substances I’m working with are 4% Paraformaldehyde and avertin. By reading about them, I learned that 4% PFA causes respiratory irritation if inhaled so heating must be performed under the chemical hood. Avertin, an anesthetic used for IP injections in mice, is good for about two weeks. When it starts to deteriorate, the pH becomes less than 5 and it is no longer effective as an anesthetic.

Knowing the facts about the substances I’m working with makes me feel more responsible for my own project and better able to discuss with others about what the necessary precautions should be when working with a certain substance. Thus, I would like to share my mentor’s advice with others. Below is the link for the OESO website; you can look up the chemicals you are working with, too!

http://www.safety.duke.edu/

I am working in the Bryan Research Building under Dr. Nicole Calakos’s guidance. You can read about the work the lab does here: http://www.neuro.duke.edu/ctn/faculty/calakos/

Today I am vibratome slicing a whole brain (we had been doing half brains) because I need to confocal image 3 different areas of the striatum, which you can’t see if you only use the half brain. A confocal microscope is pretty difficult to use when you use it for the first time. The Zeiss 410 microscope that I use is located in the French Science building. Everyone who uses it is trained because it is a delicate instrument. The confocal gives you clearer images than you would get on an epifluorescence microscope because it gives you an average of multiple layers of tissue or cells so you can see the spatial arrangement (3-D look). I look at brain slices that are 50 micrometers thick and have three fluorescent colors (red, blue, and green). The great thing about the confocal is that you can see all three at once on your slide and if you focus the image correctly (which takes me awhile because I am still getting used to it) the results are pretty neat. Here’s a picture of the confocal I use: