The 8-μm pore transwell chambers (Costar, MA, USA) were used to perform the transwell migration and invasion assays, either without (for migration assays) or with Matrigel (for invasion assays). We suspended 1.2 × 105/mL infected cells in the upper chambers with a 200-μL serum-free medium and added a 500-μL complete medium into the bottom chambers. After a 24-h incubation, we removed the cells on the upper chambers, fixed the cells on the lower compartment with 4% paraformaldehyde, and imaged them with a microscope (Olympus, Tokyo, Japan).
8 μm pore transwell chamber
Manufactured by Corning
Sourced in United States
The 8-μm-pore transwell chambers are a specialized lab equipment designed for cell culture applications. These chambers feature a porous membrane with 8-micron pores, allowing for the study of cell migration, invasion, and other cellular interactions across a defined barrier.
Automatically generated - may contain errors
Lab products found in correlation
Matrigel, by BD (11 mentions)
M4 microplate reader, by Molecular Devices (2 mentions)
Odyssey infrared imaging system, by LI COR (2 mentions)
Matrigel, by Corning (1 mentions)
Matrigel, by Solarbio (1 mentions)
Crystal violet, by Merck Group (1 mentions)
Celltiter glo reagent, by Promega (1 mentions)
Axio observer, by Zeiss (1 mentions)
Celltracker green cmfda, by Thermo Fisher Scientific (1 mentions)
Ix81 zdc inverted fluorescence microscope, by Olympus (1 mentions)
Hemacolor solution, by Merck Group (1 mentions)
Opti mem, by Thermo Fisher Scientific (1 mentions)
Ckx53, by Olympus (1 mentions)
23 protocols using 8 μm pore transwell chamber
1
Transwell Migration and Invasion Assay
2
Transwell Invasion Assay for GBC Cells
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GBC cell invasion was examined in 8-μm pore transwell chambers (Corning Costar, Cambridge, MA). Briefly, the lower surface of the transwell was coated with 10 μg gelatin and the upper side was coated with 25 μg (0.5 μg/μl) reconstituted basement membrane substance (Matrigel; BD Biosciences). Next, fresh medium containing 10% FBS was placed in the lower chamber as a chemoattractant. GBC cells were incubated for 24 h in a medium containing 1% FBS, trypsinized, and suspended at a final concentration of 1.5 × 105 cells/ml in FBS-free medium. The GBS cell suspension (100 μl) was loaded into each of the upper wells, and the chamber was incubated at 37 °C for 24 h. Cells were fixed and subjected to H&E staining. Chemotaxis activity was quantified by counting the cells that migrated to the lower side of the filter with an optical microscope. The numbers of cells in eight random fields were counted for each assay. The experiments were repeated three times.
3
Transwell Assay for Cell Migration and Invasion
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8-μm-pore transwell chambers (Corning Costar, Corning, USA) coated with Matrigel (BD, San Diego, CA, USA) were used for cell invasion assay, while those without any pre-treatment were for cell migration detection. Equal number of suspended transfected cells (1.0–1.2 × 104 cells for migration assay; 3.0–3.5 × 104 cells for invasion assay) in 200 μL FBS-free medium were loaded into the upper chamber of each 24-well transwell chamber and 600 μl 10%-FBS medium was added into each lower chamber. After cultured at 37 °C with 5% CO2 48 h, the cells on the lower surface were stained by crystal violet after the non-adhering cells in the upper chambers were scraped. Images were obtained by the inverted microscope and cell counting was performed by the software ImageJ.
4
Transwell Migration Assay Protocol
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For migration assay, 8 μm-pore transwell chambers (Corning Costar, USA) were used without Matrigel. Each well of 24-well plates was filled with the RPMI-1640 complete culture medium containing 10% FBS. Then, approximately 1 × 105 cells in 500ul FBS-free medium were pipette into the upper chamber slowly. The incubation time is 48 h for HCT116 and LOVO. After being washed by PBS 3 times, cells on the upper surface of the chamber were removed lightly using a cotton swab. Then, the cells migrated to the lower surface of the chamber were fixed using 4% paraformaldehyde (PFA). At the end of the experiment, cells were stained with 0.1% crystal violet solution and counted and analyzed by ImageJ software.
5
Transwell Assay for Cell Migration and Invasion
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A total of 1 × 105 (link)
cells suspended in serum-free culture medium were seeded in the upper compartment of 8-μm-pore transwell chambers (Corning) with 400 μL complete medium added in the lower compartment. For invasion assay, the 1:20 mixture of Matrigel (Corning) and serum-free medium was plated onto the upper compartment one hour in advance. After 24-h incubation at 37°C, cells passing through the filter were fixed and stained using crystal violet for 10 min. Images of stained cells were collected and the number of migrated or invasive cells were calculated using ImageJ. For each group, 3 and 2 replicated wells were used in migration assay and invasion assay, respectively, 4 fields per well were blindly chosen for counting.
cells suspended in serum-free culture medium were seeded in the upper compartment of 8-μm-pore transwell chambers (Corning) with 400 μL complete medium added in the lower compartment. For invasion assay, the 1:20 mixture of Matrigel (Corning) and serum-free medium was plated onto the upper compartment one hour in advance. After 24-h incubation at 37°C, cells passing through the filter were fixed and stained using crystal violet for 10 min. Images of stained cells were collected and the number of migrated or invasive cells were calculated using ImageJ. For each group, 3 and 2 replicated wells were used in migration assay and invasion assay, respectively, 4 fields per well were blindly chosen for counting.
6
Transwell-based Cell Migration and Invasion Assays
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Cell migration assays were conducted using 24-well plates with 8-μm pore Transwell chambers (Corning, NY, USA). The lower chamber was filled with culture medium containing 10% FBS. A549 or NCI-H460 cells were suspended at a density of 5 × 104 cells in MEM or 1 × 105 cells in RPMI medium, respectively, without FBS and added to the upper chamber. Three days after seeding, the cells on the bottom layer surface were stained with 0.05% crystal violet dye, and the intensity values were measured using an Odyssey infrared imaging system (LI-COR Biosciences). For the cell invasion assay, cells were seeded in the upper chamber filled with Matrigel (BD Biosciences, Erembodegem, Belgium). Five to 7 days after seeding, the cells on the bottom layer surface were stained with 0.05% crystal violet dye, and after treatment with dimethyl sulfoxide (DMSO), the absorbance was measured at 590 nm using an M4 microplate reader (Molecular Devices, CA, USA).
7
Transwell Assays for Cell Migration and Invasion
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Transwell cell migration and invasion assays were performed as described previously [25 (link)]. Briefly, cell migration assays were conducted using 24-well plates with 8-μm pore Transwell chambers (Corning, NY, USA). The lower chamber was filled with culture medium containing 10% FBS. NCI-H460 cells were suspended at a density of 5 × 104 cells/well in RPMI medium, without FBS and added to the upper chamber. Three days after seeding, the cells on the bottom layer surface were stained with 0.05% crystal violet dye, and the intensity values were measured using an Odyssey infrared imaging system (LI-COR Biosciences). For the cell invasion assay, cells were seeded at a density of 1 × 105 cells/well in the upper chamber filled with Matrigel (BD Biosciences, Erembodegem, Belgium). Five to seven days after seeding, the cells on the bottom layer surface were stained with 0.05% crystal violet dye, and after treatment with DMSO, the absorbance was measured at 590 nm using an M4 microplate reader (Molecular Devices, CA, USA).
8
Evaluating Cell Invasion Potential
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Transwell assays were performed using 8 μm-pore transwell chambers (Corning company) coated with Matrigel (BD Bioscience). 500 μl of medium containing 10% FBS (the attractant) was added into lower chambers, and 100 μl serum-free medium into upper chambers. The transfected BGC-823 cells were plated in the upper chambers. After 48-hour incubation, the cells on the bottom surface of the upper chambers were cleaned with a cotton swab. Invading cells on another surface were fixed with 4% paraformaldehyde and stained with 0.1% crystal violet for 30 min. Five random fields under a microscope were photographed and then the cell numbers were counted.
9
Transwell Cell Migration and Invasion Assay
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Cell migration and invasion assays were performed with 8-μm pore Transwell chambers (Corning, NY, USA) as previously described [23 (link)]. Overall, 5 × 104 or 8 × 104 cells in 200 μl serum-free medium were seeded in the upper chamber coated with or without Matrigel for migration or invasion assays, respectively. The lower chambers were filled with 600 μL medium with 10% FBS for migration or with 20% FBS for invasion. After 24–36 h, the cells on the bottom surface of the membrane were fixed with 95% ethanol and stained with 0.1% crystal violet. Five random fields (× 200) were counted using a microscope to assess cell migration and invasion.
10
Transwell Invasion Assay for Cell Migration
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Transwell invasion assay was performed using 8-μm-pore transwell chambers (Corning, New York, NY). Briefly, EV and PGDH(+) cells were starved for 12 h. Then, cells were trypsinized, washed twice with serum-free medium, and resuspended in serum-free medium. Cells were counted and seeded into the upper insert of the transwell chamber while RPMI-1640 medium containing 10% FBS were added to the lower chamber. Cells were then incubated at 37°C with 5% CO2. Cells that invaded through the Matrigel (Solarbio, Beijing, China)-coated membrane were trypsinized and CCK-8 reagent was added to treated cells and incubated at 37°C for 2 h. Optical density (OD) was measured at 450 nm with a microplate reader.
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