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

Journal:

Article Title: Influence of Native and Hypochlorite-Modified Low-Density Lipoprotein on Gene Expression in Human Proximal Tubular Epithelium

doi:

Figure Lengend Snippet: Presence of HOCl-modified (lipo)proteins in kidney tissues. As confirmed by immunohistology both a control human kidney (A) and a kidney with MCG (C) lack staining for HOCl-modified (lipo)proteins. In contrast (B) in renal transplant showing acute tubular damage (ATD) staining for HOCl-modified (lipo)proteins can be seen in tubular epithelia and in interstitial mononuclear cells. A: avidin-biotinylated enzyme method. B and C: APAAP method (as described in Materials and Methods); magnification ×400.

Article Snippet: This was followed by an anti-mouse biotinylated secondary antibody (PharMingen 02002D, dilution 1:250 in 1% BSA/PBS at 22°C for 1 hour).

Techniques: Modification, Staining, Avidin-Biotin Assay

Journal:

Article Title: Influence of Native and Hypochlorite-Modified Low-Density Lipoprotein on Gene Expression in Human Proximal Tubular Epithelium

doi:

Figure Lengend Snippet: Presence of HOCl-modified (lipo)proteins and MPO in kidney samples from patients with MGN. A: HOCl-modified epitopes are present in MGN with decreased renal function. Protein droplets of proximal tubules are strongly reactive. A large cytoplasmic bleb protruding into the tubular lumen (arrow) is stained. B: In the same biopsy as shown in A, staining for MPO can be seen in protein droplets of proximal tubules and in some interstitial mononuclear cells. C: In MGN with normal excretory function no staining for HOCl epitopes can be detected in tubules by immunohistology. D: Staining for MPO in MGN with regular function can be seen in isolated monocytes in the interstitium; there is some unspecific stain along peritubular capillaries, but tubules do not show positivity for MPO to a significant extent. A and C: Avidin-biotinylated enzyme method, B and D: APAAP method.

Article Snippet: This was followed by an anti-mouse biotinylated secondary antibody (PharMingen 02002D, dilution 1:250 in 1% BSA/PBS at 22°C for 1 hour).

Techniques: Modification, Staining, Isolation, Avidin-Biotin Assay