Towards Neuronal Organoids: A Method for Long-Term Culturing of High-Density Hippocampal Neurons
Figure 9
Confocal laser scanning microscope images captured from (A) 67 DIV E18 hippocampal culture at 10X magnification and (B–C) close-up view of 14 DIV E18 hippocampal culture originally at 40X magnification. (A) Culture was immunostained for IP3R (green) and β-catenin (red). Differential staining patterns suggest multiple cell types within the neurosphere. Note the localization of β-catenin within the center of the cellular mass. As a positive control for our β-catenin staining, 14 DIV cultures were treated with Wnt5 (B) and Wnt3 (C) prior to fixing and staining to re-localize β-catenin either in the cytoplasm (Wnt5) or in the nucleus (Wnt3). Notice that the β-catenin (red) is more dominant in the nucleus after treatment with Wnt3 (C) versus Wnt5 (B), demonstrating that the β-catenin staining is highly specific.