A migration model of pancreatic cancer cells was constructed using 8 μM Transwell chambers (Costar, Corning, Cambridge, MA, USA). A total of 5 × 104 cells in 200 μL of culture medium plus 0.1% FBS were planted into the upper chamber, and 700 μL of medium containing 30% FBS was added to the lower chamber. After incubation for 24 h, cells were fixed with 4% paraformaldehyde and stained with 0.5% crystal violet. Cells on the underside of the chamber membrane were observed and counted in 5 randomly chosen horizons under an inverted microscope to obtain the average value.
8 μm transwell chamber
Manufactured by Corning
Sourced in United States
The 8-μm Transwell chambers are a type of laboratory equipment designed for cell culture studies. These chambers feature a porous membrane with an 8-micrometer pore size, which allows for the study of cell migration and invasion through the membrane.
Automatically generated - may contain errors
Lab products found in correlation
Matrigel, by BD (19 mentions)
Inverted microscope, by Olympus (3 mentions)
Matrigel, by Merck Group (3 mentions)
Matrigel, by Corning (3 mentions)
Crystal violet, by Merck Group (2 mentions)
Crystal violet, by Sangon (1 mentions)
Matrigel coated membrane, by Corning (1 mentions)
Matrigel gel, by Corning (1 mentions)
Crystal violet, by Beyotime (1 mentions)
Celltracker green, by Thermo Fisher Scientific (1 mentions)
Digital sight ds u2, by Nikon (1 mentions)
Inverted microscope, by Nikon (1 mentions)
Crystal violet staining solution, by Beyotime (1 mentions)
Complete medium, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Te2000, by Nikon (1 mentions)
Am6000 microscope, by Leica (1 mentions)
Cck 8 solution, by Dojindo Laboratories (1 mentions)
Triton x 100 solution, by Merck Group (1 mentions)
51 protocols using 8 μm transwell chamber
1
Transwell Migration Assay for Pancreatic Cancer
2
Transwell Migration Assay Protocol
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The Transwell migration assay was carried out by 8μm Transwell Chambers (Corning Costar). The upper chambers were filled with 5×103 cells in 200μL media supplemented without FBS. Five hundred microliter DMEM supplemented with 10%FBS were plated in the lower chambers. After 24 hours of culture at 37°C, residual cells on the upper surface of the mem membrane were removed. The cells on the submembrane surface were fixed with 4% paraformaldehyde, and stained with crystal violet, and then take pictures under a microscope.
3
Transwell Migration and Scratch Wound Healing Assays
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For transwell migration assay, 50000 cells were seeded in 8 μm transwell chambers (Corning Costar) per well in 0.2 ml RPMI 1640 with 1% FBS. The lower chamber of the transwell device was filled with 500 μl RPMI 1640 with 10% FBS. At the indicated times, remove excess cells in the chambers using phosphate buffered saline (PBS). Cells on the lower surface of the membrane were fixed in 4% formaldehyde then stained with 1% crystal violet. Then, photographs were taken using a microscope (Olympus, IX71).
For scratch wound healing assay, cells were seeded in 6-well plates to form a confluent monolayer of 80–90% confluence in 2.0 ml RPMI 1640 with 10% FBS. Then, drawing a straight line using a 200 μl pipet tips across monolayer cell. After washing the cells three times with PBS, fresh medium with 1% FBS was added. At the indicated times, photographs were taken using a microscope (Olympus, IX71).
For scratch wound healing assay, cells were seeded in 6-well plates to form a confluent monolayer of 80–90% confluence in 2.0 ml RPMI 1640 with 10% FBS. Then, drawing a straight line using a 200 μl pipet tips across monolayer cell. After washing the cells three times with PBS, fresh medium with 1% FBS was added. At the indicated times, photographs were taken using a microscope (Olympus, IX71).
4
Transwell Assay for Monocyte Migration
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The 8‐μm Transwell chambers (Corning Costar) were used to perform the migration assay. 600 μl TPE supernatant was added to the bottom chamber. 2 × 105 purified nonclassical monocytes isolated from TPE were suspended in 100 μl RPMI 1640 medium and plated into the top chamber. After incubating at 37°C in 5% CO2 atmosphere for 3 h, the cells through the membrane to the lower membrane surface were fixed in 4% paraformaldehyde for 10 min and stained with Wright‐staining, then viewed and photographed under a digital microscope (Olympus BX51; Olympus). To investigate whether the CX3CL1‐CX3CR1 axis contributed to nonclassical monocyte migration, blocking experiments were performed by adding 1 μg/ml anti‐CX3CL1 mAb (MAB3652R‐100, R & D Systems).
5
Transwell Assay for Cell Migration and Invasion
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Cell migration was assessed in 8‐μm transwell chambers (Costar Inc., Cambridge, MA, USA) in accordance with the manufacturer's instructions. OS cells were seeded at 2.5 × 104 cells per well followed by incubation for 16 h at 37 °C. For the cell invasion assay, 1 × 105 cells were added to Matrigel (BD Biosciences, Franklin Lakes, NJ, USA) coated chambers for 48 h in accordance with the manufacturer's instructions. After incubation, cells on the upper side were scraped off by cotton swabs, and the migrated or invasive cells on the bottom of the membrane were fixed with 4% formaldehyde for 30 min. Then, cells were stained with 0.1% crystal violet staining solution. Cells were counted in four randomly selected microscopic fields and the average numbers were calculated.
6
Transwell Invasion Assay for PTC Cells
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The invasion ability of PTC cells was analyzed using 8 μm transwell chambers (Costar, Corning, NY, USA) and standard growth factor reduced (GFR) BD Matrigel (BD Biosciences, San Jose, CA, USA) [25] . PTC cells suspended in 200 μL serum-free medium were seeded to the matrigel-pre-coated upper chambers. A total of 500 μL RPMI-1640 medium added with 10% FBS was added into the lower chambers to act as the chemoattractant. After 24 h incubation, the non-invaded PTC cells on the upper surface of the membrane were removed mechanically. Invaded PTC cells on the lower surface of the membrane were fixed using methanol and stained using crystal violet (Sangon Biotech). Number of invaded PTC cells was analyzed in five random fields. Magnification time: 100×.
7
Transwell Cell Migration Assay
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The 8-μm Transwell chambers (Corning Glass Works, Corning, N.Y., USA) were coated by Matrigel (diluted at 1: 8 and not used in the migration assay). Cells were made into single cell suspension and seeded into the apical chambers at 5 × 104 cells/100 μL. The basolateral chambers were appended with 600 μL medium containing 10% FBS and the cells were incubated for 48 h. Subsequently, cells in the apical chambers were removed and the transmembrane cells were fixed with 5% glutaraldehyde and stained by 0.1% crystal violet dye for 10 min. The cells were photographed under a microscope, 5 fields of view were collected in each membrane and the transmembrane cells were counted.
8
Investigation of Anti-metastatic Effects
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Investigation of the anti-metastatic aspects of each formulation was performed by the “migration assay” in cell lines. The migration of MDA-MB-231 and SK-BR-3 cells was performed using 8 μm transwell chambers (Corning, New York, NY, USA). In this method, MDA-MB-231 and SK-BR-3 cells at a density of 5 × 104 cells per well were placed in an upper chamber with no serum medium, while the lower chamber was filled with 500 μL of medium supplemented with 20% FBS. After 48 h incubation with NiCoFe2O4@L-Silica@C-Niosome, the cells migrating to the lower surface of the membrane were stained and counted. The percentage of migrated cells was examined by counting cells under a microscope (Olympus, Tokyo, Japan) in 3 randomly selected fields [37 (link)].
9
Transwell-Based Migration and Invasion Assays
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The migration and invasion assays were performed using 8-μm Transwell chambers (Corning Life Science, Tewksbury, MA, USA). The cells (1×104 cells/well) were pretreated with BAY117082 (15 μM) and were loaded onto the upper chamber with 200 μL of serum-free RPMI 1640 medium. The lower chambers contained 500 μL of RPMI 1640 with 2.5% FBS, with or without 100 ng/mL SDF-1. The cells in the chambers were incubated for 24 h at 37°C. The cells could migrate through the membrane for the migration assay and through a matrigel-coated membrane for the invasion assay (Corning Life Science, Tewksbury, MA, USA). Nonmigrated cells were removed from the upper surface of the chamber with a wet cotton swab, and the cells on the lower surface of the chamber were stained using the Wright-Giemsa method as described previously [16 (link)]. At least 4 randomly selected fields were counted, and the average number from these 4 counts is presented.
10
Transwell Assay for Cell Migration and Invasion
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Transwell assays were performed using 8-μm Transwell chambers (Corning Incorporated, NY, U.S.A.). Transwell chambers coated with Matrigel (BD Biosciences, NJ, U.S.A.) were used for cell invasion. However, cell migration was detected using Transwell chambers without BD. An upper chamber with HEC-1-B cells (5×104/well) and a lower chamber containing a medium with 20% FBS were prepared. After 24 h, the cells were fixed and stained. Finally, migrated and invading cells were measured using an inverted microscope (Olympus Corporation, Tokyo, Japan).
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