Figure 4. Comparison of confocal and 2-photon imaging on CUBIC cleared testes.

(a) XZ planes of testis treated with CUBIC and acquired either by confocal or 2-photon microscopy, with or without laser compensation. No brightness and contrast enhancement was applied. Nuclei were stained with propidium iodide and pseudocolored. (b) Fluorescence intensity quantification. Mean fluorescence intensity was normalized and plotted against the imaging depth. All data are mean ± SEM of 3 ROI acquired on 1 testis.