Optimal Laser and Detector Filter Combinations for Fluorescent Proteins

The people at the CWRU flow cytometry core recent did a clean reinstall of one of their instruments, which meant that we had to re-set up our acquisition template. I still ended up eyeballing what would be the best laser / filter sets based on the pages over at FPbase.org, but I had a little bit of free time today, so I decided to work on a project I had been meaning to do for a while.

In short, between the downloadable fluorescent spectra at FPbase, as well as known instrument lasers and detector bandpass filters, I figured I could just write a script that essentially takes in whatever fluorescent protein with spectra that you have downloaded, and essentially makes a table showing you which laser + detector filter combinations give you the highest amount of fluorescence.

Here’s the R script on the lab GitHub page. I made it for the two flow cytometers and two sorters I use at the CWRU cytometry core, although it would presumably be pretty easy to change the script to make it applicable for whatever instruments are at your place of work. So here’s a screenshot of a compendium of the results for these instruments:

Nothing too surprising here, although it’s still nice / interesting to see the actual results. While it’s somewhat obvious since the standard Aria has no Green or Yellow-Green laser, we should not do any sorting with mCherry on it. Instead, we should use the Aria-SORP, which has the full complement of lasers we need.