Measures to reduce autofluorescence

Model Organism: Worms Methods & Reagents
1

Hi,

I’m administering Cy3 dye-labeled conjugated nanoparticles to worms, but because of the array of the conjugates on these particles, they do not show significant fluorescence in comparison to the worm’s lipofuscin autofluorescence under the Cy3 channel. I know younger worms do not have as much of this issue, but it still remains a significant impairment to my results since I have to turn up the gain and exposure time when imaging the worms to such an extent that the autofluorescence creates too much background that interferes with my imaging of the particle uptake. Are there any known methods in which gut granule autofluorescence can be drastically reduced without causing significant side effects?

2

We have had good luck with using the JJ1271 glo-1(zu391) X strain for fluorescent studies. This mutation causes a lack of gut granules in the animals but no other obvious effects. We are looking at sperm, so you should make sure they don’t have any minor defects I don’t know about that would not be compatible with your studies.

This strain is available at CGC.

Good luck,

Tim Kroft

3

I usually wash and starve the worms for a couple of hours in M9. This also reduces the autofluorescence in the gut.

Good luck,
Marlon