A day in the life of me in the lab is like a 90s sitcom, where you can predict what’s going to happen but each episode brings it’s own laughs. Each day in the lab is essentially a step in my experiment process, with it’s own victories and defeats, which keeps me looking forward to what each day brings.
First things first, I get to lab and check on my fly babies. I have around 20 vials of flies to take care of so I go in and make sure they’re alive and well. If it’s getting too crowded or if they need new food I flip the flies into new vials. I then collect whatever line of flies I need for dissection and take them to the dissection scopes. Dissection has come to be my favorite part of the experiments I do. After a lot of practice, I think I’ve gotten to a place where fly brain dissection has become far less stressful and actually enjoyable. I grab my forceps and the buffer solution and dissect the brains I need to, at a rate of about 15 flies per hour. I have to dissect a line in about an hour to prevent the brain tissue from deteriorating. I then fix the brains in a PFA solution and wash them thoroughly.
The next step in the process is antibody staining to target the DIP-alpha expression in the brains. We usually put the brains in primary antibody overnight in a 4 degree C fridge, so if this is where I am in the process on a given day, I will come in and get the brains from the fridge and wash them thoroughly to remove the primary antibody. After that I’ll put them in secondary antibody, which will ultimately give us the fluorescence we need to identify DIP-alpha expression under the microscope. This staining process occurs for 2 hours at room temperature and is followed by yet another cycle of washing.
Finally, all the dissection and staining and washing leads up to the defining part of the experiment: imaging. This is where we can see if any of our work paid off. This part involves mounting the brains neatly on a slide and using a confocal microscope to view the brains and any specific signals from the antibody staining. I take pictures of the data and use the results to determine which flies I need to dissect next, what antibody I should change, and start the process all over again.
