Don’t Throw Stones in a Glass-Bottomed Dish

We all know the old saying, “Don’t Throw Stones in a Glass House”…. Well, those same principles apply to axotomies in glass-bottomed Petri dishes. After tedious preparation of a zebra finch DRG culture in a 35mm plate, I was ready to image calcium fluctuations and Wallerian degeneration on the confocal microscope. All of the necessary ingredients had been added: Fluo-4 to stain the calcium under FITC illumination, EDTA to chelate the extracellular calcium, and FDUR/NGF to continue growth. This was going to be our first experiment in the live cell chamber with controlled CO2, temperature, humidity, and air flow. The glass bottom of the dish was allowing for clear resolution on the pre-axotomy pictures and I was ready to induce injury on the axons.

[Read more…]