The transfected cells were seeded into 24-well upper chamber (Corning, NY, USA) pre-coated with matrigel (BD, New Jersey, USA). The lower chamber was filled with RPMI-1640 medium containing 10% FBS. 24 h later, the invasive cells were stained with crystal violet (Sigma, MO, USA) for 15 min. Photos were taken under optical microscope (Olympus).
24 well upper chamber
Manufactured by Corning
Sourced in United States
The 24-well upper chamber is a component of a cell culture system designed for in vitro experiments. The upper chamber, which features 24 individual wells, is used to hold and contain cell samples or test materials during the experimental process. This product serves as a versatile tool for researchers and scientists conducting various cell-based studies and assays.
Automatically generated - may contain errors
Lab products found in correlation
Matrigel, by BD (3 mentions)
Crystal violet, by Merck Group (2 mentions)
Matrigel, by Merck Group (2 mentions)
Optical microscope, by Olympus (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Crystal violet, by Beyotime (1 mentions)
Penicillin streptomycin, by Beyotime (1 mentions)
Paraformaldehyde (pfa), by Beyotime (1 mentions)
Crystal violet solution, by Merck Group (1 mentions)
Inverted microscope, by Nikon (1 mentions)
Crystal violet staining solution, by Merck Group (1 mentions)
5 protocols using 24 well upper chamber
1
Matrigel-coated Invasion Assay
2
Glioma Cell Migration and Invasion Assay
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For migration assay, glioma cell suspensions were placed in a 24-well upper chamber (Corning, USA), while 1 % penicillin/streptomycin (Beyotime, China) and 10 % FBS (Gibco, USA) were supplemented in the lower chambers. After incubation for 24 h at 37 °C, the glioma cells were fixed with 4 % paraformaldehyde and then stained with crystal violet (0.1 %; Beyotime, China). Migrated glioma cells were counted under a microscope. For the invasion assay, Matrigel (BD, USA) was applied to the upper chamber and then initially maintained at 37 °C for half an hour.
3
Transwell Invasion Assay Protocol
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Cell invasion ability was carried out by Transwell invasion assay. Briefly, transfected cells were seeded into 24-well upper chamber (Corning, CA USA) pre-coated with Matrigel (Sigma, MO, USA). The lower chamber was filled with DMEM medium containing 10% FBS. After culturing for 24 h, the invade cells were stained with 0.1% crystal violet (Sigma) for 10 min. The number of invaded cells was counted under a light microscope (Nikon, Japan).
4
Transwell Assay for Cell Invasion
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Cell invasion ability was determined by transwell assay. Briefly, cells were seeded into 24-well upper chambers (Corning, NY, USA) pre-coated with Matrigel (Millipore, MA, USA) in serum-free medium. After 48 h, the invasive cells were fixed with 4% paraformaldehyde, followed by staining with 0.25% crystal violet solution (Sigma-Aldrich Co., St Louis, MO, USA) for 20 min. Then stained cells were observed and counted under an inverted microscope (Nikon, Japan), and five different microscopic views were selected randomly for analysis.
5
Invasion Assay for U87 and LN229 Cells
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For invasion assays, U87 and LN229 cells (5 × 104 cells/well resuspended in 200 mL MEM or DMEM) were added to the 24-well upper chambers (8-µm pore size; Corning Co., Corning, USA) with Matrigel (BD Biosciences). Next, 600 µL of DMEM containing 10% FBS was added to the bottom chamber. After 24 h of incubation, the culture medium in the upper chamber was discarded and the non-migrated cells were removed. The cells passed through the membrane were fixed with methanol and stained with a 1% crystal violet staining solution (Sigma-Aldrich). Additionally, five randomly selected fields from various angles were used for cell counting, image capturing, and observation under a 400x magnification microscope.
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