I am working from a robust protocol I use routinely and have a good workflow on the microscope so am certain the issue is the primary antibody. I have tried three different DCX antibodies (two from SYSY) and this one from Invirtogen. This is by far the best one, labelling the full cell projections nicely but is also super noisy.

I have tried different primary concentrations, HIER, different secondary concentrations, different secondaries, always use a secondary only control and have checked the tissue's autofluorescence. On the confocal I have tried changing the laser power, detector windows, dwell time and pinhole size but nothing is changing the signal:noise ratio.

I generally see very hit and miss DCX staining and am very happy to see the structures I do here, but is there something else I could be doing to better this during staining, imaging or processing after?