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

Figure 4.

Figure 4

Non-specific staining demonstrated by DCX primary omission experiment with unadsorbed or adsorbed secondary antibodies.

The sections were incubated with NeuN antibody and secondary antibodies A10029 (green, corresponding to NeuN) and ab150072 (red, Abcam) without (A) or with (B) tissue adsorption. Panels a and c in A and B are the spectral profiles for excitation wavelengths of 561 nm (upper) and 488 nm (lower). Panels b and d are the high magnification images of the white frames in the low magnification images of the cortex (panel e), indicating the test sites of the corresponding spectrograms in panels a and c of A and B. The white arrows in b of A and B indicate the contour of the neurons labeled by secondary ab150072 only. The color curves in A-a and c and B-a and c correspond to the color frames in panel A-b and d, B-b and d. Red curve: Spontaneous autofluorescence; green and blue: non-specific labeling by secondary antibody; yellow curve: background fluorescence. Scale bars: 10 µm in A-b and d, 20 µm in B-b and d, and 200 µm in A-e and B-e. DCX: Doublecortin.