please help, problems with immunostaining Dros larva brain

[Joan Sim]

Dear all,

I am facing problems with homogenous immunostaining of the whole Dosophila larva (3rd instar) brain, as I found after confocal microscopy that the staining / antibody penetration into the interior of the brain was poor. May I know what are some of the solutions to overcome this problem and which holder device should I perform this immunostaining experiment in so as to obtain good mixing during the immunostaining process and do not have to worry about the loss of the very small-sized larvae brains?

I would greatly appreciate any suggestions given. Thank you very much for your kind help.

Best regards,
Joan Sim