R. Wegerhoff et O. Breidbach, INTRACELLULAR DYE INJECTION OF PREVIOUSLY IMMUNOLABELED INSECT NEURONS IN FIXED BRAIN-SLICES, Journal of neuroscience methods, 53(1), 1994, pp. 87-93
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.