Journal: MRS bulletin

Article Title: Functional imaging of brain organoids using high-density microelectrode arrays.

doi: 10.1557/s43577-022-00282-w

Figure Lengend Snippet: Figure 2. Human cerebral organoids comprise diverse cell types relevant to the study of neuronal circuit development and physiology. (a) Immunohistochemistry (IHC) revealed the presence of mature neurons (Tau/NeuN), astrocytes (GFAP), and oligodendrocytes (OLIG2) after 100 days of differentiation; nuclei were stained with DAPI (in blue). Scale bars are 50 μm. (b) Quantification of NeuN-positive cells. The ratio of the number of NeuN-positive cells to the total number of cells was quantified for both hCO lines (3 hCOs per line, for each hCO > 5000 cells). The ratio of NeuN-positive cells to total cells did not differ significantly between the two lines. (c) Single-cell RNA sequencing (scRNA- seq) was performed to quantify the cellular composition of hCOs and their comparability. The results are summarized in two pie charts with the identified cell type fractions displayed for each line. For both lines the results confirmed the presence of dopaminergic, GABAergic, glutamatergic and cholinergic neurons, neural crest cells (NCCs), radial glia (RG), astrocytes and oligodendrocytes.

Article Snippet: The following primary and secondary antibodies were used in this study: Pax6 (BioLegend, San Diego, CA, USA, #901301, 1:300), Sox2 (Sigma-Aldrich, AB5603, 1:300), FoxG1 (Abcam, ab18259, 1:200), Tuj1 (BioLegend, #801202, 1:800), Ctip2 (Abcam, ab18465, 1:200), Tbr1 (Abcam, ab31940, 1:300), MAP2 (Thermo Scientific, PA1-10,005, 1:800), Tau (Thermo Scientific, MN1000, 1:500), NeuN (Boster Bio, Pleasanton, CA, USA, M11954-3, 1:300), GFAP (Novus Biologicals, Englewood, CO, USA, NB300-141 , 1:500), goat anti-mouse IgG (H + L) highly cross-adsorbed secondary antibody, Alexa Fluor Plus 488 (Thermo Scientific, A32723, 1:400), goat anti-rabbit IgG (H + L) highly cross-adsorbed secondary Antibody, Alexa Fluor 568 (Thermo Scientific, A11036, 1:400), goat anti-rat IgG (H + L) cross-adsorbed secondary antibody, Alexa Fluor 647 (Thermo Scientific, A21247, 1:400) and goat anti-chicken IgY (H + L) cross-adsorbed secondary antibody, Alexa Fluor Plus 647 (Thermo Scientific, A32933, 1:400).

Techniques: Immunohistochemistry, Staining, RNA Sequencing