INTRACELLULAR DYE INJECTION OF PREVIOUSLY IMMUNOLABELED INSECT NEURONS IN FIXED BRAIN-SLICES

Our method combines intracellular dye injection and immunohistochemist ry. Under optical control, Lucifer Yellow was injected into immunohist ochemically identified neurons that reside in fixed tissue. The techni que allows visualization of the complete arborization patterns of immu nostained neurons. Injections were performed on small neurons (somata < 10 mu m in diameter). The technique works on microslices of insect b rain. Standard immunohistochemical procedures have only been varied sl ightly, omitting Triton X-100 treatment. Anti-Lucifer Yellow immunohis tochemistry, or alternatively the photoconversion technique, enables e xtension of the morphological analysis of these cells to the electron microscopic level. In the present study, Lucifer Yellow injections wer e performed on immunohistochemically pretreated brain microslices (ant i-Locusta tachykinin II antiserum) of the beetle Tenebrio molitor.