haematoxylin qs  (Vector Laboratories)

Journal: bioRxiv

Article Title: Phaeochromocytomas and paragangliomas harbour tumour-initiating SOX2+ stem cells

doi: 10.1101/2025.07.03.663006

Figure Lengend Snippet: In all panels, cells immunopositive for SOX2 are in brown, cells counterstained with haematoxylin. (A) Non-metastatic phaeochromocytomas PCC1 to PCC3 with mutations in NF1. (B) Non-metastatic phaeochromocytoma PCC4 with mutation in SDHC ; (C) Non- metastatic phaeochromocytomas PCC5 to PCC7 with mutations in RET (D) Malignant phaeochromocytoma PCC8 and non-malignant paraganglioma PGL9 with mutations in VHL (E) Non- malignant paragangliomas PGL12 to PGL14 and malignant paragangliomas PGL15 to PGL17 with mutations in SDHB (F) Non-malignant phaeochromocytomas PCC10 and PCC11 with no known mutations. (G) Malignant phaeochromocytomas PCC12 and PCC13 with no known mutations. Scale bars 50µm.

Article Snippet: Nuclei were counterstained with Vector Haematoxylin QS (Vector Laboratories, CAT#H-3404-100) and slides were mounted in VectaMount Permanent Mounting Medium (Vector Laboratories, CAT#H-5000-60).

Techniques: Mutagenesis

Journal: bioRxiv

Article Title: Phaeochromocytomas and paragangliomas harbour tumour-initiating SOX2+ stem cells

doi: 10.1101/2025.07.03.663006

Figure Lengend Snippet: (A) Immunohistochemistry confirming the presence of SOX2+ cells in foetal adrenal medullae at 12 post-conception weeks (PCW), 17PCW and 19PCW. SOX2 positive cells in brown, tissues counterstained with haematoxylin. Scale bars 100µm. (B) In vitro isolation of foetal adrenomedullary cells at 19PCW leads to the generation of adherent colonies from 3 days under defined stem cell-promoting media. Scale bars 50µm. In vitro isolation of adult adrenomedullary cells from PCC30 benign phaeochromocytoma with no known mutations and PGL33, malignant paraganglioma with a germline mutation in SDHD , under defined stem cell-promoting media. Adherent cells and colonies seen at 24 and 96 hours. Scale bars 50µm. (C) Immunofluorescence using antibodies against SOX2 and TH confirms protein expression in PCC30 adherent colonies after two weeks in culture. Scale bars 50µm.

Article Snippet: Nuclei were counterstained with Vector Haematoxylin QS (Vector Laboratories, CAT#H-3404-100) and slides were mounted in VectaMount Permanent Mounting Medium (Vector Laboratories, CAT#H-5000-60).

Techniques: Immunohistochemistry, In Vitro, Isolation, Mutagenesis, Immunofluorescence, Expressing

Journal: bioRxiv

Article Title: Phaeochromocytomas and paragangliomas harbour tumour-initiating SOX2+ stem cells

doi: 10.1101/2025.07.03.663006

Figure Lengend Snippet: (A) Single-cell suspension and labelling of foetal 19PCW cells with CellTracker green CMFDA label. Scale bars 500µm. (B) Pipeline for the in vitro isolation and in ovo transplantation of human SOX2+ cells (schematic created in BioRender.com ). (C) Chick chorioallantoic membrane (CAM) xenograft of PGL33 after 4 days of incubation. Brightfield and UV fluorescence images of the dissected CAM, with a visible cell mass established on the vascularised CAM. The silicon O-ring is visible. Red and yellow boxes indicate the magnified regions shown below, depicting the mass. (D) Immunostaining on formalin-fixed, paraffin sections of the xenograft in C. Immunofluorescence using antibodies against human nuclear antigen (HNA, red)) confirming the presence of human cells in the mass. Nuclei counterstained with DAPI. Immunohistochemistry with antibodies against SOX2 (brown), confirming the presence of SOX2-expressing cells. Nuclei counterstained with haematoxylin. Scale bars 100µm. (E) Cryosections through the lungs of chicks where CAMs were successfully grafted with either foetal adrenal stem cell cultures, isolated stem cell cultures from PCC30 or isolated stem cell cultures from PGL33. Green fluorescent cells are detected in PGL33 cultures confirming metastasis. Nuclei counterstained with DAPI. Scale bars 100µm, except on right-hand panels where scale bars are 20µm. (F) Immunofluorescence staining on cryosections through chick lungs of a PGL33 xenograft with metastasis, using antibodies against SOX2 (magenta) and TH (green). Nuclei counterstained with DAPI (blue). The greyscale images of single channels are shown for signal clarity. Yellow arrows indicate SOX2 positive cells, green arrows indicate TH positive cells and white arrows indicate SOX2;TH double-positive cells. Nuclei counterstained with DAPI. Scale bars 50µm. XenoT = xenotransplanted.

Article Snippet: Nuclei were counterstained with Vector Haematoxylin QS (Vector Laboratories, CAT#H-3404-100) and slides were mounted in VectaMount Permanent Mounting Medium (Vector Laboratories, CAT#H-5000-60).

Techniques: Suspension, In Vitro, Isolation, In Ovo, Transplantation Assay, Membrane, Incubation, Fluorescence, Immunostaining, Immunofluorescence, Immunohistochemistry, Expressing, Staining