Journal: PLoS ONE

Article Title: Silver and Gold Nanoparticles Exposure to In Vitro Cultured Retina – Studies on Nanoparticle Internalization, Apoptosis, Oxidative Stress, Glial- and Microglial Activity

doi: 10.1371/journal.pone.0105359

Figure Lengend Snippet: Fluorescent immunostaining: Iba1 (green) and ED1 (red) for detecting microglial cells (DAPI, blue). In the NP-exposed retinas (20 nm Ag (G–I), 80 nm Ag (J–L), 20 nm Au (M–O), 80 nm Au (P–R)) as well as in the control retinas (A–C) the Iba1- and ED–positive cells were located in the IPL, INL and GCL in all groups (A–C). AgNO 3 -exposed retina displayed the same staining pattern, but with a stronger intensity especially for the ED1-staining (D–F). Graph shows numbers of microglial cells and data is given as mean ±SD (n = 4 explants/group). * p <0.05 compared to control. GCL = ganglion cell layer, INL = inner nuclear layer, IPL = inner plexiform layer, ONL = outer nuclear layer, OPL = outer plexiform layer. Scale bar equals 200 µm.

Article Snippet: Sections were then incubated with primary antibodies, rabbit anti-glial fibrillary acidic protein (GFAP, 1∶1500 DAKO Cytomation, Glostrup Denmark), rabbit anti- Iba1 (1∶200, WAKO, Japan), and rat anti-mouse ED1 (CD68, 1∶1000, Nordic Biosite, Sweden), 1∶1000, overnight at 4°C overnight, and thereafter detected by incubation in secondary antibodies for 2 h. Secondary antibodies included were Texas Red-conjugated donkey anti-rabbit antibody (1∶200; Abcam, Cambridge, UK), Alexa 488 goat anti-rabbit IgG (Molecular Probes) and Alexa 564 goat anti-rat (Molecular Probes).

Techniques: Immunostaining, Control, Staining

Journal: PLoS ONE

Article Title: Silver and Gold Nanoparticles Exposure to In Vitro Cultured Retina – Studies on Nanoparticle Internalization, Apoptosis, Oxidative Stress, Glial- and Microglial Activity

doi: 10.1371/journal.pone.0105359

Figure Lengend Snippet: A–D. Iba1-immunohistochemistry revealing microglial cells with ramified (A), intermediate (B), round (C) and amoeboid (D) morphology, respectively. Quantification of the fractions of microglia in three different stages of activation was performed: intermediate or “early activated” cells as judged from the morphology and co-expression of ED1 while round- and amoeboid morphologies are regarded as “late activated” cells. The graph shows the distribution of activation stages, normalized to total numbers of Iba1-positive cells/explant; values ±SD (n = 4 explants/group) are given. * p <0.05 compared to control. GCL = ganglion cell layer; INL = inner nuclear layer; ONL = outer nuclear layer. Scale bars equal 50 µm.

Article Snippet: Sections were then incubated with primary antibodies, rabbit anti-glial fibrillary acidic protein (GFAP, 1∶1500 DAKO Cytomation, Glostrup Denmark), rabbit anti- Iba1 (1∶200, WAKO, Japan), and rat anti-mouse ED1 (CD68, 1∶1000, Nordic Biosite, Sweden), 1∶1000, overnight at 4°C overnight, and thereafter detected by incubation in secondary antibodies for 2 h. Secondary antibodies included were Texas Red-conjugated donkey anti-rabbit antibody (1∶200; Abcam, Cambridge, UK), Alexa 488 goat anti-rabbit IgG (Molecular Probes) and Alexa 564 goat anti-rat (Molecular Probes).

Techniques: Immunohistochemistry, Activation Assay, Expressing, Control

Journal: BMC Cancer

Article Title: Local iron homeostasis in the breast ductal carcinoma microenvironment

doi: 10.1186/s12885-016-2228-y

Figure Lengend Snippet: FPN1-expressing leukocytes in carcinomas are predominantly M2-like. Sections of normal breast tissue, DCIS (ductal carcinoma in situ) and IDC (invasive ductal carcinoma) to reveal the presence of cells of the macrophage lineage (CD68), and its classical polarization phenotypes, M1-like (CD80) and M2-like (CD163), in FPN1-expressing leukocyte infiltrate. For details see Materials and Methods (Original magnification × 100- upper CD68 images, ×400- squared image series)

Article Snippet: Immunohistochemical staining was performed in 2 μm-thick TMA sections with the following antibodies: rabbit polyclonal anti-human hepcidin-25 antibody (dilution 1:500, Abcam, Cambridge, UK [ ]), rabbit polyclonal anti-human ferroportin 1 antibody (FPN—1:500, Novus Biologicals Europe, Cambridge, UK [ ]), rabbit polyclonal anti-human ferritin antibody (FT—1:1000, Sigma-Aldrich, MO, USA [ ]), mouse monoclonal anti-human CD71 (TFR1 [clone 10 F11]- 1:80, Novocastra, Newcastle, UK [ ]), mouse monoclonal anti-human CD68 (clone Kp-1, 1:2000, A. Menarini Diagnostics, CA, USA), mouse monoclonal anti-human CD163 (clone MRQ-26, 1:100, Cell Marque, CA, USA), mouse monoclonal anti-human CD80 (37711, 1:100, R&D Systems, MN, USA), rabbit polyclonal anti-human CD4 (clone H-370, 1:250, Santa Cruz Biotechnology, TX, USA) and mouse monoclonal anti-human CD8 (clone C8/144B, 1:100, Cell Marque, CA, USA).

Techniques: Expressing, In Situ

Journal: BMC Cancer

Article Title: Local iron homeostasis in the breast ductal carcinoma microenvironment

doi: 10.1186/s12885-016-2228-y

Figure Lengend Snippet: Representative images of the FPN1 analysis by Imaging Flow Cytometry in breast cancer core biopsies. Epithelial cells (EC) are stained by an anti-cytokeratin (CK) FITC. T lymphocytes (T Ly) were identifiable by CD3 PerCP-Cy5.5, B lymphocytes (B Ly) by CD20 PE-Cy7 and macrophages (M0) by CD68 PE-Cy7 (Original magnification × 400)

Article Snippet: Immunohistochemical staining was performed in 2 μm-thick TMA sections with the following antibodies: rabbit polyclonal anti-human hepcidin-25 antibody (dilution 1:500, Abcam, Cambridge, UK [ ]), rabbit polyclonal anti-human ferroportin 1 antibody (FPN—1:500, Novus Biologicals Europe, Cambridge, UK [ ]), rabbit polyclonal anti-human ferritin antibody (FT—1:1000, Sigma-Aldrich, MO, USA [ ]), mouse monoclonal anti-human CD71 (TFR1 [clone 10 F11]- 1:80, Novocastra, Newcastle, UK [ ]), mouse monoclonal anti-human CD68 (clone Kp-1, 1:2000, A. Menarini Diagnostics, CA, USA), mouse monoclonal anti-human CD163 (clone MRQ-26, 1:100, Cell Marque, CA, USA), mouse monoclonal anti-human CD80 (37711, 1:100, R&D Systems, MN, USA), rabbit polyclonal anti-human CD4 (clone H-370, 1:250, Santa Cruz Biotechnology, TX, USA) and mouse monoclonal anti-human CD8 (clone C8/144B, 1:100, Cell Marque, CA, USA).

Techniques: Imaging, Flow Cytometry, Staining