Journal: PLoS ONE

Article Title: Interactions between Spider Silk and Cells – NIH/3T3 Fibroblasts Seeded on Miniature Weaving Frames

doi: 10.1371/journal.pone.0012032

Figure Lengend Snippet: A: SEM of weaving frame with silk, fibroblasts and extracellular matrix, note the dense central regions; magnitude ×39, scale bar represents 100 µm. B: Live/Dead staining of fibroblasts on silk, assembled in crosslink pattern, viable cells are stained green, dead cells (and spider silk via autofluorescence) red; magnitude ×100, scale bar represents 50 µm. C: Immunofluorescence of spider silk, fibroblasts, and extracellular matrix, Ethidium bromide for staining of the cell nuclei in orange , collagen I antibody in blue , autofluorescence of spider silk green ; magnitude ×40, scale bar represents 200 µm. D: Immunofluorescence of spider silk, fibroblasts, and extracellular matrix, DAPI staining of the cell nuclei in blue , fibronectin antibody in pink , spider silk not visible due to use of secondary antibodies beyond autofluorescence spectrum; magnitude ×40, scale bar represents 200 µm.

Article Snippet: For controls, cover slips were either rinsed with 70% Ethanol, autoclaved and coated with 0.15 mg/ml fibronectin solution in PBS (Biochrom AG, Berlin, Germany) for 30 minutes at room temperature (RT) and washed with PBS, or were covered with 1 mg/ml collagen A (Biochrom AG) solution for 30 minutes at 37°C and washed with PBS, respectively.

Techniques: Staining, Immunofluorescence

Journal: PLoS ONE

Article Title: Interactions between Spider Silk and Cells – NIH/3T3 Fibroblasts Seeded on Miniature Weaving Frames

doi: 10.1371/journal.pone.0012032

Figure Lengend Snippet: A: Cell count of the proliferation assay, numbers of visible DAPI-stained cell nuclei were counted in fields of vision (FOV) with n = 10 for each specimen, with each specimen treated in quadruplicate. 1 = Collagen-coated cover slip, 2 = Fibronectin-coated cover slip, 3 = weaving frame with native spider silk, 4 = weaving frame with trypsin-treated spider silk. Error bars indicate standard error means, asterisks mark significance level p<0.05. B: Measurement of relative fluorescence of Tubulin in cells either on spider silk weaving frames, collagen-coated controls or fibronectin-coated controls, depicted as relative fluorescent units (RFU). Error bars indicate standard error means, asterisks mark significance level p<0.05. C: Mitotic index of proliferating cells either on spider silk weaving frames, collagen-coated controls or fibronectin-coated controls, calculated by dividing RFU values of Tubulin-positive cells by RFU values of Phospho-Histone H3-positive cells (see text). Error bars indicate standard error means, asterisks mark significance level p<0.05.

Article Snippet: For controls, cover slips were either rinsed with 70% Ethanol, autoclaved and coated with 0.15 mg/ml fibronectin solution in PBS (Biochrom AG, Berlin, Germany) for 30 minutes at room temperature (RT) and washed with PBS, or were covered with 1 mg/ml collagen A (Biochrom AG) solution for 30 minutes at 37°C and washed with PBS, respectively.

Techniques: Cell Counting, Proliferation Assay, Staining, Fluorescence

Journal: PLoS ONE

Article Title: Interactions between Spider Silk and Cells – NIH/3T3 Fibroblasts Seeded on Miniature Weaving Frames

doi: 10.1371/journal.pone.0012032

Figure Lengend Snippet: Immunofluorescence scan of scaffolds seeded with cells on days 1, 2, and 5, stained with Tubulin antibody, columns are spider silk, Collagen control and Fibronectin control; scale bar represents 10 mm.

Article Snippet: For controls, cover slips were either rinsed with 70% Ethanol, autoclaved and coated with 0.15 mg/ml fibronectin solution in PBS (Biochrom AG, Berlin, Germany) for 30 minutes at room temperature (RT) and washed with PBS, or were covered with 1 mg/ml collagen A (Biochrom AG) solution for 30 minutes at 37°C and washed with PBS, respectively.

Techniques: Immunofluorescence, Staining, Control