Fig.Ā 2From: A method for estimating relative changes in the synaptic density in Drosophila central nervous systemMilder fixation, longer permeabilization, and incubation with antibody gave optimum Bruchpilot staining. Squash preparation of VNC of Drosophila 3rd larval instar. aāc Bruchpilot staining with different combinations of incubation periods for post-fixation and antibody (Ab) incubation with 0.3% PT-X permeabilization for 10Ā min. d Milder post-fixation for 10Ā min with 10Ā min 0.3% PT-X permeabilization. e 20Ā min 0.3% PT-X permeabilization with milder post-fixation for 10 and 60Ā min Ab incubation. f 20Ā min 0.3% PT-X permeabilization and 30Ā min 0.1% PBT-X blocking with milder post-fixation for 10 and 60Ā min Ab incubation. Magnification Ć40 oil objective, N.A.ā=ā1.3; Scale bars: 25Ā Āµm. The experiments were performed using a set of 5ā10 isolated VNCs, and the images represent the majority observation amongst each setBack to article page