Sonication-facilitated Immunofluorescence Staining of Late-stage Embryonic and Larval Drosophila Tissues In Situ

The overall goal of this procedure is to visualize or sate tissues in C two using fluorescence microscopy following sonication. This is accomplished by first collecting embryonic and larval samples and then fixing them using formaldehyde, heptane, and methanol. The second step is to sonicate the fixed sample in order to remove the cuticle to permit antibody penetration.

Next, the sample is immuno stain with primary and fluorescent secondary antibodies. The final step is to mount the sample in a glycerol based Antifa solution. Ultimately, confocal or epi fluorescent microscopy is used to visualize the developing tissue as well as protein expression and localization.