Modified Boyden chambers with polycarbonate‐membrane inserts (pore size, 8 μm; Falcon; Becton Dickinson) coated with 30 μg of Matrigel (BD) were used for cell invasion assays.41 Cells were suspended in medium containing 10% NuSerum (Thermo) and placed in the upper chambers, while 1 mL of medium was placed in each of the lower chambers. After 12 h incubation, cells were fixed with methanol and stained with 10% Giemsa stain solution (Sigma). Cell numbers were counted under a microscope at ×100 magnification. Experiments were performed at least twice, and each sample was assayed in triplicate.
Boyden chambers with polycarbonate membrane inserts
Manufactured by BD
Boyden chambers with polycarbonate-membrane inserts are a type of laboratory equipment used for cell migration and invasion assays. They consist of a two-compartment system separated by a polycarbonate membrane. Cells are seeded in the upper compartment, and their ability to migrate or invade through the membrane into the lower compartment is measured.
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2 protocols using boyden chambers with polycarbonate membrane inserts
1
Matrigel-Coated Boyden Chamber Assay
2
Matrigel-Coated Cell Invasion Assay
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Modified Boyden chambers with polycarbonate-membrane inserts (pore size, 8 μm; Falcon; Becton Dickinson, Franklin Lakes, NJ) coated with 12 μg of Matrigel (BD; San Jose, CA) were used for cell invasion assays. Stable transfectants were suspended in culture medium containing 10% NuSerum (Invitrogen). Cells (2.5 × 104) were placed in the upper chambers, and 1 ml of medium was placed in the lower chambers. After incubation for 24 h at 37 °C, the cells were fixed with methanol for 10 min at room temperature and then stained for 30 mins at room temperature with a 50 μg/ml solution of propidium iodide (PI) (Sigma). The number of cells on each membrane was counted under a microscope at a magnification of 50× using the Analytical Imaging Station software package (Imaging Research Inc., St. Catharines, ON, Canada).
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