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: PLoS ONE

Article Title: Anti-Tumor and Immune Enhancing Activities of Rice Bran Gramisterol on Acute Myelogenous Leukemia

doi: 10.1371/journal.pone.0146869

Figure Lengend Snippet: (A), (B) MTT assay of WEHI-3 cells treated with different concentrations of RBDS for 24, 48 and 72 h, and gramisterol for 24 h. Percent cell viability was calculated relative to the untreated control. The experiments were performed in triplicate and data are presented as mean±SEM. Significant difference was considered at p <0.05 compared with the control. (C) Phase contrast photographs and DAPI-DNA staining of WEHI-3 cells after treatment with RBDS 0, 50, 100 and 200 μg/mL for 48 h. The apoptotic bodies are indicated by white arrowheads. (D) Gel electrophoresis of DNA extracted from RBDS treated-WEHI-3 for 48 h. DNA ladder or fragmentation indicated apoptosis event in the cells. (E) Flow cytometry analysis of WEHI-3 cells after 48 h RBDS-treatment and annexin-V staining. Percent cell count of apoptosis detection was demonstrated from the average of two experiments.

Article Snippet: The treated cells were cold fixed with methanol, air dried and stained with DAPI nuclear staining dye (Boehringer, Ingelheim, Germany) at 37°C for 30 min. Apoptosis hallmarks such as cell shrinkage, membrane bleb, and chromatin condensation were examined under a fluorescence microscope.

Techniques: MTT Assay, Control, Staining, Nucleic Acid Electrophoresis, Flow Cytometry, Cell Counting

Journal: Oncology Letters

Article Title: Application of a novel liquid biopsy in patients with hepatocellular carcinoma undergoing liver transplantation

doi: 10.3892/ol.2018.8019

Figure Lengend Snippet: Identification of CTCs in HCC patients using iFISH platform. CK: Green, CEP8: Orange, DAPI: Blue, CD45: Red. (A) CK+/CD45-/DAPI+/CEP8=2; (B) CK+/CD45-/DAPI+/CEP8>2; (C) CK-/CD45+/DAPI+/CEP8>2. (D) CTC cluster. (E) CK-/CD45+/DAPI+/CEP8=2, WBC. iFISH, immunostaining-fluorescence in situ hybridization; CTCs, circulating tumor cells; HCC, hepatocellular carcinoma; CK, cytokeratin; CEP8, centromere probe 8; CD, cluster of differentiation; WBC, white blood cell.

Article Snippet: Immunofluorescence reagent (cat no. 7900001; Menarini Silicon Biosystems, Inc.) was used according to the manufacturer's protocol and included anti-CK (CK8, CK18 and CK19; 0.0006%) antibody conjugated to phycoerythrin, anti-CD45 antibody (0.0012%) conjugated to allophycocyanin and nuclear dye DAPI (0.005%).

Techniques: Immunostaining, Fluorescence, In Situ Hybridization