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: 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: After washing three times with PBS, sections were incubated with PureBlu DAPI (Bio-Rad, Hercules, CA, USA, #1351303) for 3 min and mounted with ProLong Gold antifade mounting medium (Thermo Scientific, #P36930).

Techniques: Immunohistochemistry, Staining, RNA Sequencing

Journal: International Journal of Molecular Sciences

Article Title: Cyclin B3 Deficiency Impairs Germline Stem Cell Maintenance and Its Overexpression Delays Cystoblast Differentiation in Drosophila Ovary

doi: 10.3390/ijms19010298

Figure Lengend Snippet: cycB3 is required intrinsically for controlling the fate of GSCs. ( A , B ) Ovaries were stained with anti-Hts antibody (red) to visualize fusomes, 4',6-diamidino-2-phenylindole (DAPI) dye to label nuclei (blue), and anti-GFP antibody (green) to show the cycB3 expression pattern; ( A’ , A ) Ovary from an Oregon fly (wild-type). GSCs (indicated by dashed circles) were GFP negative (noted by arrowheads); ( B’ , B ) Ovary carrying a transgene P{ cycB3P-cycB3-gfp }. GSCs (indicated by dashed circles) were GFP positive (green, noted by arrowheads); ( C – G ) Fourteen-day-old germaria stained with anti-Vasa antibody (green, germ cells), and with anti-Hts antibody (red, spectrosomes and fusomes). GSCs are indicated by arrows. ( C ) cycB3 mutant germarium with one GSC; ( D ) cycB3 mutant with empty germarium. ( E , F ) cycB3 mutant ovaries were rescued by the transgenes of nos-gal4:vp16 ; UASp-cycB3 and nosP-cycB3 . Germarium with two GSCs. ( G ) cycB3 mutant flies, carrying the genotype, c587-gal4 ; UASp-cycB3 . Ovary with an empty germarium. Scale bars: 5 μm.

Article Snippet: DAPI dye (Yeasen, Shanghai, China) was used to visualize cellular nuclei.

Techniques: Staining, Expressing, Mutagenesis

Journal: International Journal of Molecular Sciences

Article Title: Cyclin B3 Deficiency Impairs Germline Stem Cell Maintenance and Its Overexpression Delays Cystoblast Differentiation in Drosophila Ovary

doi: 10.3390/ijms19010298

Figure Lengend Snippet: The mutation in cycB3 fails to affect oocyte formation. ( A – D ) The deficiency of cycB3 fails to lead to apoptosis in GSCs. Germaria from wild-type ( A ) and cycB3 2 mutants ( B ) were labeled by TUNEL (red, indicated by arrowheads) and stained with anti-Vasa antibody (green) and DAPI dye (blue); Germaria from FRT controls ( C ) and FRT cycB3 2 mutants ( D ) were labeled by TUNEL (red, indicated by arrowheads), stained with anti-GFP antibody (green) and DAPI (blue); ( E – H ) The cycB3 mutation in CB fails to impair its differentiation into oocytes; Germaria from wild-type ( E ) and cycB3 2 mutants ( F ) were stained with anti-Orb antibody (red, indicated by arrowheads), anti-Vasa antibody (green) and DAPI (blue). The outlines of germaria are drawn by dashed circles. Germaria from FRT controls ( G ) and FRT cycB3 2 mutants ( H ) were stained by anti-Orb antibody (red, indicated by arrowheads), anti-GFP antibody (green) and DAPI (blue). GSCs are noted by arrows ( A , B , E , F ); GSC clones are indicated by dashed circles and cyst clones are noted by solid circles ( C , D , G , H ). The outlines of germaria are drawn with dashed circles ( A – F ). Scale bars: 5 μm.

Article Snippet: DAPI dye (Yeasen, Shanghai, China) was used to visualize cellular nuclei.

Techniques: Mutagenesis, Labeling, TUNEL Assay, Staining, Clone Assay

Journal: International Journal of Molecular Sciences

Article Title: Cyclin B3 Deficiency Impairs Germline Stem Cell Maintenance and Its Overexpression Delays Cystoblast Differentiation in Drosophila Ovary

doi: 10.3390/ijms19010298

Figure Lengend Snippet: Over-expression of cycB3 notably increases the number of CBs. Ovaries from wild-type ( A ) and cycB3 -overexpression flies ( B – D ) were stained with anti-Hts antibody (red), anti-Vasa antibody (green), and DAPI dye (blue). The micrographs were stacked together in the Z-axis direction, to visualize spectrosome-containing GSCs (indicated by arrows) and CBs (noted by arrowheads). Scale bars: 5 μm.

Article Snippet: DAPI dye (Yeasen, Shanghai, China) was used to visualize cellular nuclei.

Techniques: Over Expression, Staining