Imaging, Imaging and Analysis

A typical day in the Calakos lab starts with reading through the literature on the topic of habitual behavior and goal-directed behavior. Sometimes I start by listening in on the lab meetings which are once every week in the morning. These days I have finally obtained permission to go work with Zeiss 880 inverted confocal microscope, so I have started to spend half of my day imaging mouse brain slices, and striatum in particular.

Obtaining the correct settings for each of the four lasers (I am trying to image with four fluorophores simultaneously) takes a surprising amount of time. It is critical for me to use the same settings for each experiment for a fair comparison. Furthermore, taking the images themselves takes hours. For example, one 20x tile scan with three fluorophores and 10 by 16 frames takes around 1 hour and 30 minutes. A whole brain constitutes of many individual slices, which will have to be imaged separately. I have a lot to do in the future.

In the last weeks of the program, I am planning to analyze these images in order to figure out the changes in ensembles of neurons activated for habitual behavior vs goal-directed behavior, and compare them with afferents into the different areas of dorsal striatum. I imagine this will be plenty (and more) work to keep me occupied in the coming weeks.

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