Cell invasion was detected using a 24-well transwell chamber (Corning, New York, USA) with coated with Matrigel (BD Biosciences). Cell migration was detected using a 24-well transwell chamber (Corning, New York, USA). Briefly, SW1353 and U2OS cells transfected with indicated mimics in 100 μL serum-free medium were plated in the top chamber. Next, 600 μL complete medium was added to the lower chambers. 48 h later, the invaded and migrated cells were counted according to the manufacturer's instructions [18 (link)].
24 well transwell chamber
Manufactured by Corning
Sourced in United States, China, United Kingdom
The 24-well transwell chambers are a laboratory equipment used for cell culture studies. They consist of a multi-well plate with a semi-permeable membrane insert that allows for the co-culture of cells in separate compartments while facilitating the exchange of soluble factors between them.
Automatically generated - may contain errors
Lab products found in correlation
Matrigel, by BD (143 mentions)
Matrigel, by Corning (55 mentions)
Crystal violet, by Merck Group (19 mentions)
Inverted microscope, by Olympus (19 mentions)
Crystal violet, by Beyotime (15 mentions)
Crystal violet, by Sangon (11 mentions)
Crystal violet, by Solarbio (9 mentions)
Matrigel matrix, by BD (9 mentions)
Matrigel, by Solarbio (9 mentions)
Inverted light microscope, by Olympus (7 mentions)
Matrigel, by Merck Group (7 mentions)
Paraformaldehyde (pfa), by Merck Group (7 mentions)
Matrigel solution, by BD (5 mentions)
Methanol, by Sangon (5 mentions)
Matrigel, by Thermo Fisher Scientific (5 mentions)
Matrigel matrix, by Corning (4 mentions)
Methanol, by Merck Group (4 mentions)
Matrigel basement membrane matrix, by BD (4 mentions)
A microscope, by Olympus (4 mentions)
Diff quik stain set, by Siemens (4 mentions)
589 protocols using 24 well transwell chamber
1
Cell Invasion and Migration Assay
2
Quantifying Cell Migration and Invasion
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Cell migration was determined using a 24-well transwell chamber (Corning, New York, USA), and cell invasion using a 24-well transwell chamber (Corning, New York, USA) coated with 50mg/L Matrigel (BD Biosciences). Briefly, indicated-mimics transfected HeLa and SiHa cells in 100 μL serum-free RPMI 1640 medium (Gibco by Life Technologies, USA) were plated in the top chamber, with a final density of 2×105 cell/well. Next, 600 μL complete medium was added to the lower chambers. Twenty-four hours later, the invaded or migrated cells were fixed with methanol, stained with crystal violet.22 (link) The images in five random fields were taken was by an Olympus BX53 microscope carrying DP72 camera (Olympus, Japan) and cell number was counted by Image J software (NIH). Parallel experiments were carried out five times independently.
3
Cell Migration and Invasion Assay
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About 5 × 104 cells in 150 μl of 1640 medium without FBS were added to the upper compartment of a 24-well transwell chamber (pore size 8.0 μm, diameter 6.5 mm, Costar, USA), and 350 μl of 1640 medium with 20% FBS was added to the lower compartment. After incubating for 9.5 hours with 5% CO2 at 37°C, the upper chamber was washed three times with PBS, and the cells were subsequently stained and fixed with 0.1% crystal violet diluted with methanol for 2 hours. After being washed three times with PBS and dried, the cells were photographed with a microscope at 200× magnification. Finally, Image-Pro Plus 6.0 software was used to count the numbers of cells in four randomly chosen areas. For invasion assay, all the procedures were the same as those described above; however, the 24-well transwell chambers were coated with 40 μl of Matrigel (at a dilution of 1 to 8) on the upward-facing side of the polycarbonate membrane, 1.5 × 105 cells were added to upper chamber, and the culture time was prolonged to 48 hours.
4
Cell Invasion and Migration Assay
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The cell invasion assay was performed using a 24-well Transwell chamber (Costar, USA) without Matrigel coating. For migration assay, a total of 2 × 105 transfected cells were added to the upper chamber of Transwell assay inserts. Then, the inserts were added to the bottom chamber wells filled with conditioned medium. After the incubation for 24 hours and stained with hematoxylin for 10 minutes, we calculated the number of cell in five random fields for each chamber. For invasion assay, transfected cells were plated in the top chamber with a Matrigel-coated membrane in the bottom chambers.
5
Cell Invasion Assay Using Matrigel
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Cell invasion assay was performed using a 24-well transwell chamber with 8 μm pores (Costar, Cambridge, MA, USA). Inserts were coated with 20 μL of Matrigel (1:3 dilution; BD Biosciences, San Jose, CA, USA). At 48 h after transfection, cells were trypsinized, and 3×105 cells in 100 μL of serum-free medium were transferred to the upper Matrigel chamber for 18 h. Media supplemented with 10% FBS were added to the lower chamber as a chemoattractant. After incubation, cells that passed through the filter were fixed with 4% paraformaldehyde and stained with hematoxylin. Invasive cells were microscopically counted in 10 randomly selected high-power fields.
6
Scratch Wound Healing Assay
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Cells were cultured into six well plates, and an artificial wound was made using a 100 μl pipette tip after 24 hours. The images of wounds were captured at 0 hour or 24 hours under an inverted microscope. In invasion assay, a 24-well Transwell chamber (Costar, USA) precoated with Matrigel (BD) was used to measure cell invasion. 0.1 ml cells suspended (3×104) were seeded into the upper chambers and maintained with serum-free medium. Lower chambers were filled with medium containing with 20% FBS. After 24 hours, the invading cells were fixed with 4% paraformaldehyde and stained by 1% crystal violet. The invading cells were counted in five randomly selected fields.
7
Quantifying Cell Invasion Using Transwell Assay
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A total of 5 × 104 transfected cells were plated into the upper chamber of a 24-well transwell chamber (Corning Costar) and covered with a matrix (200 µL per well). After culturing for 12 h in serum-free medium, the chamber was removed from the plate. The chamber was then fixed with 4% paraformaldehyde and stained with 0.5% crystal violet for 30 min. The invading cells were photographed using a microscope and analyzed using ImageJ.
8
Transwell Assay for Migration and Invasion
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To examine migration and invasion, cells were seeded into a 24-well transwell chamber (Costar, USA) with or without precoated Matrigel. Cells in the upper chamber were cultured using 200 μL serum-free medium. The lower chamber was fixed with 10% FBS-supplemented medium. After incubating for 24 h, the cells that migrated or invaded into the lower membrane surface were fixed with 4% paraformaldehyde and stained with 1% crystal violet. Cell numbers were finally counted.
9
Invasive Cell Migration Assay
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Invasive activity was estimated using [3H]thymidine-labeled cells, as previously described.9 (link) The upper surfaces of 8-μm pore size polycarbonate nucleopore filter inserts in a 24-well Transwell chamber (Corning Costar, Cambridge, MA, USA) were coated with Matrigel (30 μg/well; Becton Dickinson, Lincoln Park, NJ, USA). The [3H]thymidine-labeled cells (5 × 104 cells) were seeded into the upper part of the Matrigel-coated filters, and medium was added to the lower part. The radioactivity of the cells that invaded through the Matrigel into the lower part of the chamber was counted using a LS6500 liquid scintillation counter with a liquid scintillation cocktail (Beckman Coulter, Fullerton, CA, USA). Results are expressed as changes in invasion relative to controls.
10
Cell Migration Assay with Transwell
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A 24-well Transwell chamber (8 μm aperture; Corning Costar, USA) was prepared overnight at 4°C with or without 100μL matrix gel substrate provided by BD Biosciences, San Jose, CA, USA. A 200ul of cell suspension containing 1×105cells/mL was inoculated into Transwell cells with or without matrix glue, and a culture medium (800 ul) containing 10% FBS was poured into the lower chamber. After 24h culture, cell fixation was done using 4% paraformaldehyde at room temperature for 20 minutes and staining was performed for 5 minutes with 0.5% crystal violet dye. The cell count was recorded afterwards.
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