Journal: European Journal of Histochemistry : EJH

Article Title: Spatio-temporal course of macrophage-like cell accumulation after experimental embolic stroke depending on treatment with tissue plasminogen activator and its combination with hyperbaric oxygenation

doi: 10.4081/ejh.2012.e14

Figure Lengend Snippet: Triple fluorescence labelling of CD68, glial and neuronal markers.

Article Snippet: Next, the sections were extensively rinsed with TBS and processed with highly purified biotinylated goat-anti-mouse IgG (Dianova, Hamburg, Germany; 1:500 in TBS containing 2% bovine serum albumin = TBS-BSA) for one h. Following several rinses with TBS, the sections were applied to preformed streptavidin/biotinyl-peroxidase complexes (12.5+5 µg/mL TBS-BSA) and then stained with diaminobenzidine/ammonium nickel sulphate as chromogen according to Härtig et al.

Techniques: Fluorescence

Journal: European Journal of Histochemistry : EJH

Article Title: Spatio-temporal course of macrophage-like cell accumulation after experimental embolic stroke depending on treatment with tissue plasminogen activator and its combination with hyperbaric oxygenation

doi: 10.4081/ejh.2012.e14

Figure Lengend Snippet: Triple fluorescence labelling of markers for macrophages and microglia, 4 weeks after embolic stroke in the right middle cerebral artery territory. (A) The staining of CD68 with FITC-conjugated monoclonal mouse antibodies was enhanced by Cy2-tagged anti-FITC, and combined with (B) the immunodetection of CD11b based on biotinylated mouse antibodies visualized by Cy3-streptavidin, and (C) Cy5-immunolabelling of ionized calcium binding adaptor molecule 1 (Iba) in microglial cells. The merge of the three staining patterns reveals a microglial population co-expressing CD11b and Iba (purple, arrows), but in the centre also cells exist, displaying all three markers (white, arrowheads). Scale bar: 50 µm.

Article Snippet: Next, the sections were extensively rinsed with TBS and processed with highly purified biotinylated goat-anti-mouse IgG (Dianova, Hamburg, Germany; 1:500 in TBS containing 2% bovine serum albumin = TBS-BSA) for one h. Following several rinses with TBS, the sections were applied to preformed streptavidin/biotinyl-peroxidase complexes (12.5+5 µg/mL TBS-BSA) and then stained with diaminobenzidine/ammonium nickel sulphate as chromogen according to Härtig et al.

Techniques: Fluorescence, Staining, Immunodetection, Binding Assay, Expressing

Journal: European Journal of Histochemistry : EJH

Article Title: Spatio-temporal course of macrophage-like cell accumulation after experimental embolic stroke depending on treatment with tissue plasminogen activator and its combination with hyperbaric oxygenation

doi: 10.4081/ejh.2012.e14

Figure Lengend Snippet: Concomitant immunofluorescence staining of CD68 in macrophage-like cells, ionized calcium binding adaptor molecule 1 (Iba) in microglial cells and glial fibrillary acidic protein (GFAP) in astroglia 4 weeks after embolic stroke in the border zone of the infarct. (A) CD68-immunoreactivity addressed with biotinylated monoclonal mouse antibodies and visualised by Cy2-conjugated streptavidin, and (B) Iba revealed by Cy5-immunolabelling were counterstained in (C) with Cy3-coupled mouse-anti-GFAP. By merging of staining patterns in (D) activated micro- and astroglia were also elucidated around the centre of the infarct with infiltrated CD68-immunopositive macrophages. Scale bar: 50 µm.

Article Snippet: Next, the sections were extensively rinsed with TBS and processed with highly purified biotinylated goat-anti-mouse IgG (Dianova, Hamburg, Germany; 1:500 in TBS containing 2% bovine serum albumin = TBS-BSA) for one h. Following several rinses with TBS, the sections were applied to preformed streptavidin/biotinyl-peroxidase complexes (12.5+5 µg/mL TBS-BSA) and then stained with diaminobenzidine/ammonium nickel sulphate as chromogen according to Härtig et al.

Techniques: Immunofluorescence, Staining, Binding Assay