Ibidi 2-well inserts (Ibidi) with a cell-free gap were used in 12-well plates to evaluate cell migration by wound healing assay. U373 and SNB19 transfected cells were plated in triplicate, at an initial density of 70 000 and 60 000 cells in each side of the insert, respectively, and left to adhere overnight. After that, inserts were removed (0 h time-point) and wells were washed with PBS to remove dead cells and debris. The migration of the cells into the gap was imaged (2 images/well) over hours until wound closure was achieved (images were captured at the same locations over time). The gap size (pixels) was measured using an automated software (beWound - Cell Migration Tool, v1.7, BESURG, Portugal; www.besurg.com ) and manually verified and corrected when deemed necessary. At least 15 lines equally spaced across the image and perpendicular to the gap main axis were used for increased measurement representativeness.
2 well insert
Manufactured by Ibidi
Sourced in Germany
The 2-well inserts are a type of cell culture laboratory equipment designed to create a two-chamber environment for various experimental setups. These inserts are suitable for a range of cell-based assays and co-culture studies.
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Lab products found in correlation
6 protocols using 2 well insert
1
Evaluating Cell Migration via Wound Healing
2
Astrocyte Migration Assay Protocol
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Astrocytes, after stimulation, were cultured in Transwell chamber inserts with 8-μm filters (Corning, New York, USA). The upper chamber was filled with 2 × 105 cells/mL in serum-free medium, and the lower chamber was filled with 500 μL 10% DMEM. After 24 h culture, unmigrated astrocytes on the upper chamber of the membrane were eliminated and migrated astrocytes on the lower surface were fixed with 4% paraformaldehyde for 15 min and stained with 0.1% crystal violet for 20 min. The scratch wound assay was performed by culturing the treated astrocytes in 2-well inserts (Ibidi, Martin Reid, Germany) in 24-well plates (Corning, New York, NY, USA). The cells migrating into the gap were counted at 24 h.
3
Cell Migration Assay with GBML Cells
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GBML18 and GBML42 cells were platted at high density (6 × 104 cells in each compartment) in 2‐well inserts (ibidi GmbH, Gräfelfing, Germany) and incubated overnight to allow cells’ adhesion. Inserts were removed on the following day, and migration capacity was registered in photographs taken in an inverted microscope (objective lens magnification 4×; Olympus CKX41 microscope, Olympus, Tokyo, Japan). For each time point, two images/areas were recorded per well. Wound widths (in pixels) were calculated using the bewound software (version 1.7, BESURG, Braga, Portugal), and wound closures were determined in percentages.
4
Endothelial Cell Migration Assay
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A 24 well flat-bottom tissue culture plate (VWR) was coated with RTC1 and allowed to incubate for at least 4 hours. The wells were washed with PBS twice and then were allowed to dry until the 2 well inserts (ibidi) can adhere. 70 μL of cell suspension at 80,000 cells/mL were added to each side of the insert and 300 μL of EGM2 was added to the surround well. The plate was incubated overnight. The inserts were removed, the media was aspirated, and 500 μL of EGM2 was added to the well. The cells were imaged using the Lionheart FX Automated Microscope (BioTek Instruments) every hour for 24 hours. The area calculations were based on a protocol developed by BioTek Instruments.
5
Wound Healing Assay with Drug Treatment
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Cells were seeded into a 2-well insert (Ibidi, Munich, Germany) on a 6-well plate and allowed to attach overnight. A wound was created by removing the insert, and the cell-free gap was around 500 μm. Cells were allowed to migrate in the presence or absence of drugs, and images were captured at the indicated times by EVOS XL Core Cell Imaging System (Thermo Scientific).
6
In Vitro Wound Healing Assay
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Thirty thousand cells per well were seeded in the ibidi 2‐well insert (ibidi, Fitchburg, WI, USA) that was placed in a 24‐well plate and after 48 h incubation, the insert was removed, leaving a 500 μm gap between the two separate cell monolayers. Afterwards, a 24 h time‐lapse video was recorded with a time interval of 1 h. Each image was analyzed by imagej software to measure the area occupied by cells and wound area, which were used to calculate % wound area and % healed area.
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