THP-1 monocytes were pre-infected T. gondii for 4 h (MOI=4) and then allowed to adhere onto 50 μg/mL fibronectin-coated glass coverslip dishes (MatTek) for 30 min. Samples were then gently rinsed with PBS and fixed for 10 min with ice-cold 4% formaldehyde in PBS. Cells were permeabilized with 0.05% saponin and stained with monoclonal antibodies to LFA-1 (TS2/4) or VLA-4 (9F10), followed by Alexa fluorophore-conjugated secondary antibodies. Samples were imaged in PBS with the earlier-described TIRF microscopy system and a 100x oil Plan-Apochromat (N.A. 1.46) objective. Tandem Z-stack series of the same cells were captured with the earlier-described spinning disc system to visualize cells as infected or uninfected. TIRF images were analyzed in Volocity software. Cell outlines were manually traced, and integrin foci were identified using the Volocity Spot Finder function.
Fibronectin coated glass coverslip dishes
Manufactured by MatTek
Fibronectin-coated glass coverslip dishes are a type of lab equipment. They consist of glass coverslips that have been coated with the protein fibronectin. Fibronectin is a component of the extracellular matrix and facilitates cell attachment and growth.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using fibronectin coated glass coverslip dishes
1
Visualizing Integrin Dynamics in T. gondii-Infected Monocytes
2
Visualizing Integrin Dynamics in T. gondii-Infected Monocytes
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THP-1 monocytes were pre-infected T. gondii for 4 h (MOI=4) and then allowed to adhere onto 50 μg/mL fibronectin-coated glass coverslip dishes (MatTek) for 30 min. Samples were then gently rinsed with PBS and fixed for 10 min with ice-cold 4% formaldehyde in PBS. Cells were permeabilized with 0.05% saponin and stained with monoclonal antibodies to LFA-1 (TS2/4) or VLA-4 (9F10), followed by Alexa fluorophore-conjugated secondary antibodies. Samples were imaged in PBS with the earlier-described TIRF microscopy system and a 100x oil Plan-Apochromat (N.A. 1.46) objective. Tandem Z-stack series of the same cells were captured with the earlier-described spinning disc system to visualize cells as infected or uninfected. TIRF images were analyzed in Volocity software. Cell outlines were manually traced, and integrin foci were identified using the Volocity Spot Finder function.
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