Please excuse the poorly placed John Mayer reference in the title, but it was too good of an opportunity to pass up. Although the irony of how well some of the lyrics from that song fits working with fluorochromes is startling. Case and point: "Who knows how long, how long, how long she can go before she burns away." An apt reminder of the ever present possibility of quenching looming overhead and its ticking clock. But enough about John Mayer, and more about how cool it was getting to try my hand at some confocal imaging! This morning we were viewing glioblastoma cells we treated with various combination of Mito and LysoTracker stains, DAPI, Phalloidin, and antibodies against β-actin and tubulin. The confocal, by far, has the most bells and whistles of the microscopes we have used thus far this semester. Not only can you tune the lasers to fit extremely narrow excitation and emission windows, but you can also create a max intensity image that combines all the layers taken in a z-stack. The confocal took everything I thought I knew about light microscopy and blew it out the water. It was downright amazing getting to see how different laser lines and light wavelength completely alter what you see under the scope (or should I say on the computer screen). Just seeing the bright, blue nuclei light up from the DAPI stain, let alone observing where the red actin and green tubulin overlap when running those lasers together, was incredible. However, there is certainly an art to deciding which combination of antibodies and fluorochromes should be used at what wavelength to achieve your specific research goal. While I'm no artist, I certainly do enjoy (and can manage) this color-by-number science.