BEAS-2B cells were seeded on coverslips and incubated overnight in DMEM plus 10% FBS at 37°C with 5% CO2 in a humidified cell incubator. Cells were fixed using 4% paraformaldehyde and permeabilized with 0.1% Triton X-100 in phosphate-buffered saline (PBS). Cells were incubated with anti-XB13 (mouse IgG, homemade) and anti–Tks5 SH3#1 (rabbit IgG; EMD Millipore), primary antibody and secondary antibody conjugated with Alexa Fluor 594 (Molecular Probes, Eugene, OR), or Oregon green 488 (Molecular Probes). Actin was stained using CytoPainter phalloidin-iFluor 405 reagent (Abcam). Coverslips were mounted onto glass slides with Dako fluorescence mounting medium (Dako, Mississauga, Canada). Images were obtained using an Olympus FluoView Confocal FV1000-ASW and analyzed by Olympus FluoView FV10-ASW (Olympus, Tokyo, Japan) and ImageJ (National Institutes of Health, Bethesda, MD).
Fluoview confocal fv1000 asw
Manufactured by Olympus
Sourced in Japan
The FluoView Confocal FV1000-ASW is a laser scanning confocal microscope system designed for advanced fluorescence imaging. It provides high-resolution, three-dimensional imaging capabilities for a wide range of biological samples.
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2 protocols using fluoview confocal fv1000 asw
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Immunofluorescence Staining of BEAS-2B Cells
2
Immunofluorescence Imaging of Cytoskeletal Proteins
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BEAS-2B cells were seeded on coverslips and incubated overnight in DMEM + 10% FBS at 37°C with 5% CO2 in a humidified cell incubator. Cells were fixed using 4% paraformaldehyde and permeabilized with 0.1% Triton X-100 in PBS. Cells were incubated with anti-XB130 (mouse IgG, homemade), anti-Tks5 SH3#1 (rabbit IgG, EMD Millipore), anti-N-WASP (rabbit IgG, Santa Cruz) or anti-WAVE2 (rabbit IgG, Santa Cruz) primary antibody and secondary antibody conjugated with Alexa Fluor 594 (Molecular Probes) or Oregon Green 488 (Molecular Probes). Actin was stained using CytoPainter Phalloidin- iFluor 405 reagent (Abcam, Cambridge, UK). Coverslips were mounted onto glass slides with DAKO fluorescence mounting medium (Dako, Mississauga, Canada). Images were obtained using an Olympus FluoView Confocal, FV1000-ASW and analysed by Olympus FluoView FV10-ASW and ImageJ/FIJI Coloc 2 plugin (National Institutes of Health).
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