Journal: International Journal of Molecular Sciences

Article Title: Disseminated Tumor Cells (DTCs) in Patients with Cervical Cancer Reveal Mesenchymal Properties and Potential Therapeutic Targets—A New Perspective?

doi: 10.3390/ijms27114875

Figure Lengend Snippet: Positive and negative control cells for both marker panels. ( A ) HeLa (white arrows): in the first panel positive for cytokeratin (CK), Vimentin (Vim), VEGF and in the second panel positive for p16INK4A, negative for PD-L1 and CD45. ( B ) T98G (red arrows): positive for Vim. ( C ) CaSki (yellow arrows): positive for CK, Vim, VEGF, PD-L1 and p16INK4A. ( D ) MCF-7 (green arrows): positive for CK. All cell lines were negative for CD45. Only hematopoietic cells showed CD45 positive staining. Scale bar = 10 μm. Images were processed using Zeiss ZEN 3.7 software and the figure was assembled using Microsoft PowerPoint.

Article Snippet: In the first round of staining, anti-human Pan-Cytokeratin APC-conjugated REAdye_lease antibody (Cat. No. 130-123-091, Miltenyi Biotech, Bergisch Gladbach, Germany), anti-human vimentin FITC-conjugated REAdye_lease antibody (Cat. No. 130-127-022, Miltenyi Biotech, Bergisch Gladbach, Germany) and anti-human VEGF PE-conjugated REAdye_lease antibody (Cat. No. 130-118-061, Miltenyi Biotech, Bergisch Gladbach, Germany) were applied.

Techniques: Negative Control, Marker, Staining, Software

Journal: International Journal of Molecular Sciences

Article Title: Disseminated Tumor Cells (DTCs) in Patients with Cervical Cancer Reveal Mesenchymal Properties and Potential Therapeutic Targets—A New Perspective?

doi: 10.3390/ijms27114875

Figure Lengend Snippet: Representative images of patient-derived DTCs with three different profiles. ( A ) DTC positive for Vim, VEGF and p16, negative for CK, PD-L1 and CD45. ( B ) CK and Vim positive while negative for VEGF, PD-L1, p16 and CD45. ( C ) DTC positive for Vim and PD-L1, negative for all other markers. Scale bar = 10 μm. Images were processed using Zeiss ZEN 3.7 software and figure was assembled using Microsoft PowerPoint.

Article Snippet: In the first round of staining, anti-human Pan-Cytokeratin APC-conjugated REAdye_lease antibody (Cat. No. 130-123-091, Miltenyi Biotech, Bergisch Gladbach, Germany), anti-human vimentin FITC-conjugated REAdye_lease antibody (Cat. No. 130-127-022, Miltenyi Biotech, Bergisch Gladbach, Germany) and anti-human VEGF PE-conjugated REAdye_lease antibody (Cat. No. 130-118-061, Miltenyi Biotech, Bergisch Gladbach, Germany) were applied.

Techniques: Derivative Assay, Software

Journal: International Journal of Molecular Sciences

Article Title: Disseminated Tumor Cells (DTCs) in Patients with Cervical Cancer Reveal Mesenchymal Properties and Potential Therapeutic Targets—A New Perspective?

doi: 10.3390/ijms27114875

Figure Lengend Snippet: Proportion of DTC Phenotypes. The majority of detected DTCs (70%, n = 174) was positive for Vimentin without CK expression, hence revealing a mesenchymal phenotype. An epithelial phenotype was found in 13% ( n = 31) of the DTCs and 15% ( n = 37) presented epithelial–mesenchymal transition (EMT)-like properties. Chart generated using Microsoft Excel.

Article Snippet: In the first round of staining, anti-human Pan-Cytokeratin APC-conjugated REAdye_lease antibody (Cat. No. 130-123-091, Miltenyi Biotech, Bergisch Gladbach, Germany), anti-human vimentin FITC-conjugated REAdye_lease antibody (Cat. No. 130-127-022, Miltenyi Biotech, Bergisch Gladbach, Germany) and anti-human VEGF PE-conjugated REAdye_lease antibody (Cat. No. 130-118-061, Miltenyi Biotech, Bergisch Gladbach, Germany) were applied.

Techniques: Expressing, Generated

Journal: International Journal of Molecular Sciences

Article Title: Disseminated Tumor Cells (DTCs) in Patients with Cervical Cancer Reveal Mesenchymal Properties and Potential Therapeutic Targets—A New Perspective?

doi: 10.3390/ijms27114875

Figure Lengend Snippet: Sequential multi-parameter immunofluorescent staining. After applying the first antibody panel (CK, Vim and VEGF), the slides were scanned and DTC profiles were assessed. Subsequently, releasable fluorochrome-conjugates were digested and the slides were subjected to the second antibody panel (PD-L1, p16, CD45) followed by scanning and DTC detection. Created with BioRender.com.

Article Snippet: In the first round of staining, anti-human Pan-Cytokeratin APC-conjugated REAdye_lease antibody (Cat. No. 130-123-091, Miltenyi Biotech, Bergisch Gladbach, Germany), anti-human vimentin FITC-conjugated REAdye_lease antibody (Cat. No. 130-127-022, Miltenyi Biotech, Bergisch Gladbach, Germany) and anti-human VEGF PE-conjugated REAdye_lease antibody (Cat. No. 130-118-061, Miltenyi Biotech, Bergisch Gladbach, Germany) were applied.

Techniques: Staining

Journal: OncoTargets and therapy

Article Title: An isolated orbital mass with proptosis and diplopia: an uncommon metastasis from intracranial olfactory neuroblastoma in a chinese patient

doi: 10.2147/OTT.SI09929

Figure Lengend Snippet: Immunohistochemical staining of the mass. Notes: The tumor cells were positive for ( A ) SYN; original magnification: ×400; a reaction in the cytoplasm of the neoplastic cells; ( B ) CGA; original magnification: ×400; a reaction in the cytoplasm of the neoplastic cells; (C) CD56 (cluster of differentiation 56); original magnification: ×400; a strong and heavy membrane-type staining; ( D ) CK; original magnification: ×400; a reaction in the cytoplasm of the neoplastic cells; ( E ) VIM; original magnification: x400; a reaction in the cytoplasm of the neoplastic cells; ( F ) S-100 protein; original magnification: ×400; a sustentacular reaction. The tumor cells were negative for EMA and CD99 (data not shown). Abbreviations: CD, cluster of differentiation; CGA, chromogranin; CK, cytokeratin; EMA, epithelial membrane antigen; SYN, synaptophysin; VIM, vimentin.

Article Snippet: Immunohistochemical analyses for synaptophysin (SYN) (rabbit monoclonal antibody, OriGene, Rockville, MD, USA), chromogranin A (CGA; mouse monoclonal antibody; OriGene), cluster of differentiation (CD56; mouse monoclonal antibody; OriGene), cytokeratin (CK; mouse monoclonal antibody; OriGene), vimentin (VIM; mouse monoclonal antibody; OriGene), S-100 protein (mouse monoclonal antibody, OriGene), epithelial membrane antigen (EMA; mouse monoclonal antibody; OriGene), and cluster of differentiation 99 (CD99; mouse monoclonal antibody; OriGene) were performed on the paraffin sections.

Techniques: Immunohistochemical staining, Staining, Membrane

Journal: Oncology letters

Article Title: Impact of tumoral carbonic anhydrase IX and Ki-67 expression on survival in oral squamous cell carcinoma patients.

doi: 10.3892/ol.2017.6829

Figure Lengend Snippet: Figure 1. Fluorescent immunohistochemical staining imaged with an Aperio ScanScope FL® and analyzed using AQUAnalysis® software. Indicated are repre sentative fluorescence immunohistochemistry images of (A and B) CAIX and (C and D) Ki‑67 staining in OSCC samples. (A) Positive and negative controls for CAIX involved the staining of normal OSCE with/and without the CAIX antibody, respectively. (C) Similarly, positive and negative controls for Ki‑67 involved the staining of tonsil tissue with or without the Ki‑67 antibody, respectively. Representative images of OSCC samples stained for (B) CAIX and (D) Ki‑67 are indicated with examples of high and low expressers of these proteins. Merged images are pseudo‑colored blue for DAPI, green for pan‑cytokeratin, red for the biomarker of interest and grayscale for vimentin. OSCE, oral squamous cell epithelium; OSCC, oral squamous cell carcinoma; AQUA, Automated Quantitative Analysis™; CAIX, carbonic anhydrase IX; s, stromal.

Article Snippet: Briefly, the TMA slides were stained on a DAKO AutoStainer Link 48 using antibodies against vimentin (mouse monoclonal; dilution, 1:5,000; catalog no. M0725; Dako; Agilent Technologies, Inc., Santa Clara, CA, USA), CAIX (rabbit polyclonal; dilution, 1:5,000; catalog no. ab15086; Abcam, Cambridge, MA, USA) and pan-cytokeratin (PCK; guinea pig polyclonal; dilution, 1:100; catalog no. BP5069; OriGene Technologies, Inc., Herford, Germany).

Techniques: Immunohistochemical staining, Staining, Software, Fluorescence, Immunohistochemistry, Biomarker Discovery