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. 2020 Jan 9;15(7):1290–1299. doi: 10.4103/1673-5374.272610

Figure 1.

Figure 1

Distribution of DCX-positive labeling in adjacent adult monkey brain cortex sections labeled with different primary antibodies.

(A) DCX staining using different antibody combinations. Antibodies Ab18723-1, Ab18723-2, CST-4604 and SC-271390 and their corresponding fluorescein-conjugated secondaries were used on adjacent sections of the cortex. Panels a-d show brain sections stained with different antibody combinations. In each row, DCX (a’–d’), Neun (a’’–d’’) and their merged channels are presented sequentially. The photomicrographs are of layers III (panel a) and V (panels b–d) of the cortex. Yellow arrows in panel c show the fluorescence clusters, and white arrows show the contour of the labeled cells. All DCX signals are co-localized with NeuN, a mature neuron marker. Large differences in the distribution of DCX-positive cells were observed in the cortex of adjacent sections. Scale bars: 50 μm in A-a, A-b and A-d; 20 μm in A-c. (B) Emission wavelength and intensity of DCX immunofluorescence: The adjacent sections were double immunolabeled by the combination of DCX antibody (ab18723-1) and the fluorescent secondary antibody (A10036, Thermo Fisher Scientific) and the combination of NeuN antibody (ab104224) with secondary antibody (A11029), as red and green, respectively. In panel a, the low magnification image shows the localization of the detected neuron in the cerebral cortex (white frame), and the strong fluorescent point in the perikarya is indicated with the yellow arrow. Panels b and c respectively show the wavelength, intensity and localization of fluorescence in the tested points (color frames) on the pyramidal neuron immunolabeled by DCX (b’) and NeuN (c’) with an excitation wavelength of 579 nm (upper) or 495 nm. Panels b” and c” display the detected data of the emission wavelength and the fluorescence intensity for the different tested points. I–V: Different layers of the cerebral cortex. Scale bars: 200 µm in panel B-a and 50 µm in panels B-b’ and B-c’. CST: Cell Signaling Technology; DCX: doublecortin.