SFA Tech Tip – Flies, fluorophores, and filter sets

Posted On: Thursday, September 25, 2014

The following is a communication from Dr. Gregory Beitel, Northwestern University, discussing his application of the SFA filter sets with two lines of transgenic Drosophila. We have added the notes in brackets for clarity.

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“We got the red filter and it works nicely for what we are trying to do:

Bloomington stock 51323 has a dsRED and a GFP that express in the Drosophila eye using the “P3” enhancer. With the RFP filter [our GR set – green excitation plus red barrier filter] you strongly see red eyes, and with the basic GFP filter (not the bandpass) you see whitish/pink glowing eyes with both dsRED and the GFP coming through. I imagine in the bandpass filter [the recently introduced RB-GO option] the red would be much reduced, but I don’t have the bandpass filter to test.

Bloomington stock 51324 only has the eye GFP, and does not show at all in the dsRED filter. In the GFP filter the eyes are clearly GFP green. Even with just the GFP filter, it is dead easy to score which flies have both the dsRED and GFP, and which flies have only the GFP. But I’m going to keep the dsRED filter since it works well, and I imagine it is more sensitive than the GFP set looking at RFP. It just that the P3-dsRED is so strong you can see it in the GFP channel too.

By the way these flies would make excellent demo flies since they are so bright that it is scary. You can easily see them (in fact you can’t miss them) in the vial without having to put them to sleep and put them on the pad.

Thanks for creating such a great system.”