Journal: The Scientific World Journal

Article Title: Culture and Characterization of Microglia from the Adult Murine Retina

doi: 10.1155/2014/894368

Figure Lengend Snippet: Immunofluorescent staining of cultured retinal microglial cells. Microglial cells were transferred into a 16-well chamber slide and stained for (a) isotype control (insert, phase contrast image to show the cells), (b) F4/80, (c) CD11b, (d) CD45, (e) MHC-II (insert, phase contrast), (f) CD86 (insert, phase contrast), and (g) CD40 (insert, phase contrast). The samples were analysed by confocal microscopy. In all images blue indicates allophycocyanin (APC). Over 98% of the cells were positive for CD11b, CD45, and F4/80 surface markers but the staining of CD45 was much weaker compared to CD11b and F4/80. Cells were negative for MHC-II, CD86, and CD40. Data were representative of 2 experiments.

Article Snippet: The antibodies used include purified rat anti-mouse CD45 (1 : 50, BD Biosciences, Oxford, UK), biotin conjugated anti-mouse CD11b (1 : 100, AbD Serotec, Kidlington, UK), biotin conjugated anti-mouse F4/80 (1 : 100, AbD Serotec), biotin conjugated anti-mouse CD40 (1 : 100, BD Biosciences), phycoerythrin (PE) conjugated anti-mouse CD86 (1 : 100, BD Biosciences), and fluorescein isothiocyanate (FITC) conjugated anti-mouse MHC-II (BD Biosciences).

Techniques: Staining, Cell Culture, Confocal Microscopy

Journal: The Scientific World Journal

Article Title: Culture and Characterization of Microglia from the Adult Murine Retina

doi: 10.1155/2014/894368

Figure Lengend Snippet: Immunofluorescent staining of retinal microglial cells. (a) Microglial cells were transferred into a 16-well chamber slide and stimulated with LPS in the absence of L929 supernatant for 24 h. After stimulation, cells were stained for activation markers, including MHC-II (FITC), CD86 (PE), and CD40 (APC). The slides were observed by confocal microscopy. (b) Microglial cells were transferred into a 16-well chamber slide, incubated with POS-FITC at the ratio of cells to POS-FITC 1 : 5 for 18 h, and stained for CD11b (APC). (c) Microglial cells (1 × 10 4 /well) were transferred into a 96-well plate and stimulated with 1 μ g/mL of LPS for 24 h. The supernatant was collected and measured for the presence of IL-1 β , IL-12, IL-10, CCL2, GM-CSF, IL-6, TNF- α , and CCL5 using CBA. * P ≤ 0.05 compared with control nonstimulated cell supernatant. Mean ± SEM, n = 3.

Article Snippet: The antibodies used include purified rat anti-mouse CD45 (1 : 50, BD Biosciences, Oxford, UK), biotin conjugated anti-mouse CD11b (1 : 100, AbD Serotec, Kidlington, UK), biotin conjugated anti-mouse F4/80 (1 : 100, AbD Serotec), biotin conjugated anti-mouse CD40 (1 : 100, BD Biosciences), phycoerythrin (PE) conjugated anti-mouse CD86 (1 : 100, BD Biosciences), and fluorescein isothiocyanate (FITC) conjugated anti-mouse MHC-II (BD Biosciences).

Techniques: Staining, Activation Assay, Confocal Microscopy, Incubation

Journal: Journal of Clinical Immunology

Article Title: Dendritic and T Cell Response to Influenza is Normal in the Patients with X-Linked Agammaglobulinemia

doi: 10.1007/s10875-011-9639-y

Figure Lengend Snippet: The differentiation and maturation of imDC in XLA patients upon influenza virus stimulation. Monocyte-derived imDC from XLA patients ( n = 12) and normal controls ( n = 23) were treated with heat killed H1N1 (MOI = 1) for 24 h. Surface expressions of MHC-I, MHC-II, CD83, and CD86 were examined by flow cytometry. a Histogram showed the representative data of the increase in phenotype expressions after virus stimulation; b bar chart showed the mean fluorescent intensity of the phenotype expression. Data represented as mean ± SEM. * p < 0.05; ** p < 0.01

Article Snippet: After 24 h of culture with virus, imDC were collected and analyzed by immunofluorescent staining using mAbs PE/Cy5 anti-major histocompatibility complex (MHC)-I, APC anti-CD83, and PE anti-CD86 (Biolegend, San Diego, CA, USA), fluorescein isothiocyanate (FITC) anti-MHC-II (BD Pharmingen, San Jose, CA, USA) to determine phenotypic maturation with flow cytometer.

Techniques: Derivative Assay, Flow Cytometry, Expressing