Iontophoretic application of unconjugated cholera toxin B subunit (CTb) combined with immunohistochemistry of neurochemical substances: a method for transmitter identification of retrogradely labeled neurons
Luppi P.H., Fort P., Jouvet M.
Brain Res. 534 (1-2) pages : 209-224 (1990)


Materials and Methods

Materials and Methods


(A) Injection sites

(B) Retrograde labeling

(C) Artefactual labeling due to uptake by fibers of passage

(D) Anterograde tracing

(E) Double immunostaining technique



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Figure 1

Click on photomicrographs A,B,C or D to get it bigger

A: overview of a frontal section showing an unconjugated cholera toxin B subunit (CTb) injection site in the ventrolateral part of the periaqueduetal gray in a eat with 72 h of survival including 48 h of colchicine treatment (case C116).
Bar = 2 mm.

Note that at this magnification the extent of the site is overestimated when compared with D. For this reason we used the same enlargement in Figs. 1A, 2A and 3A in order to directly compare the size of the different injection sites.

B,D: photomicrographs of adjacent sections showing the same CTb injection site stained either with the biotinylated swine anti-goat IgG and the ABC-HRP complex (B) or the biotinylated donkey anti-goat IgG and the streptavidin-HRP (D). Note that the aspect but not the size of the site changes according to the system of detection.

C: photomicrograph showing the extent of necrosis in the center of the pressure injection site in a counterstained section in the proximity of that shown in B and D.

Bars in B,C,D = 500 µm.

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