The assay was done by using chambers with polycarbonate filters (pore size, 8μm) (Becton Dickinson Labware). ESCC cells were harvested and 5×104 cells in 200μL of 0.1% serum medium were placed in the upper chamber. The lower chamber was filled with 10% fetal bovine serum medium (600 μL). After 24h incubation and removal of the cells on the upper chamber of the filter with a cotton swab, the cells on the underside were fixed with 4% paraformaldehyde, stained with 0.1% crystal violet in 20% ethanol, and counted in five randomly selected fields under phase contrast microscope. The migrated cells were monitored by photographing at 400× magnification with LEICA Microscope. The assays were performed in triplicate.
Chambers with polycarbonate filters
Manufactured by BD
Chambers with polycarbonate filters are laboratory equipment designed to provide a controlled environment for various applications. These chambers have a polycarbonate material construction and feature integrated filters that help maintain a clean, isolated workspace.
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Lab products found in correlation
3 protocols using chambers with polycarbonate filters
1
Transwell Assay for Cell Migration
2
Transwell Assay for Cell Migration and Invasion
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The transwell assay for migration was performed using chambers with polycarbonate filters (pore size 8μm) (Becton Dickinson Labware). The transwell assay for invasion was performed using chambers with Matrigel. Briefly, we harvested SW579 and TPC-1 cells and placed 5 × 104 transfected cells in the upper chamber of the filter. After 24 h of incubation and removal of the cells in the upper chamber using a cotton swab, the cells on the underside were fixed with 4% paraformaldehyde, stained with 0.1% crystal violet in 20% ethanol, and counted in five randomly selected fields under a phase-contrast microscope. The migrated or invasive cells were monitored by photographing at 400× magnification using a LEICA microscope. The assays were performed in triplicate.
3
Transwell Assay for Cell Migration and Invasion
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For migration, a Transwell assay was performed using chambers with polycarbonate filters (pore size, 8 µm) (Becton Dickinson Labware). For invasion, a Transwell assay was performed using chambers with Matrigel and polycarbonate filters (pore size, 8 µm). First, U251MG and SHG-44 cells were harvested 24 h after miRNA transfection, and 5×104 transfected cells were added to 200 µL of 0.1% serum medium in the upper chamber. The lower chamber was filled with 10% FBS medium (600 µL). After 24 h of incubation and removal of the cells from the upper chamber of the filter using a cotton swab, the cells on the underside were fixed with 4% paraformaldehyde, stained with 0.1% crystal violet in 20% ethanol, and counted in five randomly selected fields under a phase-contrast microscope. The migrated or invasive cells were monitored by photographing at 200× magnification with a LEICA microscope. The assays were performed in triplicate.
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