HCT 116 and RKO cells (5 × 104 cells/well) were inoculated into a 96-well dish for culturing until cell confluence reached 90% and scratched on the cell layer using 96 wounding replicators (VP scientific). At the same time, the medium was substituted with the 1% FBS contained fresh medium. Photographs of the wound were captured at pre-set time points (0 h, 24 h, and 48 h). Finally, the percentage of migration was evaluated utilizing Image Pro Plus.
96 wounding replicator
Manufactured by V&P Scientific
Sourced in United States
The 96-Wounding Replicator is a laboratory instrument designed for the automated creation of consistent wounds or perforations in cellular monolayers or tissues. It features a 96-well format and precisely controlled wounding parameters to enable high-throughput experimentation.
Automatically generated - may contain errors
Lab products found in correlation
Celigo, by Revvity (3 mentions)
Cellomics, by Thermo Fisher Scientific (2 mentions)
24 well transwell chamber, by Corning (1 mentions)
Image pro plus software, by Media Cybernetics (1 mentions)
Transwell chamber, by Corning (1 mentions)
Celigo image cytometer, by Revvity (1 mentions)
Celigo software, by Revvity (1 mentions)
35 protocols using 96 wounding replicator
1
Wound Healing Assay in HCT 116 and RKO Cells
2
Wound Healing Assay in Lentiviral-Infected Cells
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Lentivirus-infected HCT116 and RKO cells (5 × 104 cells/well) were seeded into 96-well plates. When the confluence of the cells reached 90%, the medium was substituted with a low-concentration serum medium and scratched on the cell layer using 96 wounding replicators (VP Scientific). Then, the cells were washed two to three times with serum-free medium and incubated in an incubator with 5% CO2 at 37°C. Photographs of the wound were captured by a fluorescence microscope at 0, 24, and 48 h. Finally, the migration area was analyzed with Cellomics.
3
Wound Healing Assay in Cancer Cells
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Briefly, lentivirus infected HO-8910 and OVCAR-3 cells (5 × 104 cells/well) were seeded into a 96-well dish for 24 h culturing. When the cells confluence reached 90%, cell scratches across the cell layer were made with a 96 wounding replicator (VP scientific). The scratches were gently rinsed with PBS for 2-3 times. Photographs were taken at 8 h and 24 h using a fluorescence microscope (OLYMPUS).
4
Quantifying Cell Migration Dynamics
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The cell migration assay was performed in 24-well transwell chambers (Corning, Shanghai, China) according to the manufacturer's instructions. For the wound-healing assay, cells were seeded into 96-well plates at a density of 5 ×104 per well and cultured overnight. Wounds were created with a 96 Wounding Replicator (VP Scientific, San Diego, CA, USA). The cells were washed gently with culture medium and further cultured with medium containing 1% FBS. Images were acquired at different time points with a microscope at 100x magnification. The gap distance was evaluated using Adobe Photoshop CS6 (v13.0).
5
Wound Healing Assay with Lentivirus
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Lentivirus transfected cells were seeded at 3 × 104 cells per well onto a 96-well dish. Scratches were made by a 96-wounding replicator (VP scientific) across the cell layers while the cell confluence reached over 90%. Serum-free medium was used to rinse gently 2–3 times and floating cells were washed. RPMI-1640 medium with 0.5% FBS was added and cultured for 24 h and photographs were taken by a fluorescence microscope at 8 and 24 h. Cell migration rate of each group was calculated.
6
Wound Healing Assay in 96-well Plates
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The cells were inoculated into 96-well plates (~3×104 cells/well) and cultured in DMEM containing 10% FBS at 37°C and 5% CO2 for 24 h. After achieving an appropriate cell attachment, scrape wounds were generated with a 96 Wounding Replicator (V&P Scientific, Inc.). The cells were washed and subsequently cultured with low-serum medium (0.5% FBS). The wound healing process was then captured by microscopy using an inverted fluorescence microscope (IX73; Olympus Corporation) at intervals of 0, 24, and 48 h/0, 4, and 8 h. The percentage of cell migration was calculated utilizing Image-Pro Plus software (version 6.0; Media Cybernetics, Inc.).
7
Cell migration quantified by scratch assay
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Cell migration was measured by scratch wound healing assays. HCT116 and RKO cells infected with shCtrl or shHSPA4 lentivirus were plated into 96-well plates (5×104 cells/well) and cultured until cell confluence reached 90%. Subsequently, cell scratches were made across the cell layer by a 96-wounding replicator (VP scientific) and the cell layers gently washed. Photographs were taken using epifluorescence microscopy at 24 h and 48 h post scratching and the migration rates calculated.
8
Cell Migration Assay Protocol
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100 µL HCCC-9810 or HUCCT1 cell suspension was added to a 96-well plate (50,000 cells/well). After 24 h, the medium was replaced with low-concentration FBS (0.5%) medium, and then a 96 Wounding Replicator (VP scientific) was used to nudge the lower central part of the well to form a scratch. After washing with FBS-free medium, medium containing 0.5% FBS was added and scanned with Cellomics (Thermo), which was recorded as 0 h. Cells were cultured at 37 °C in an incubator with 5% CO2. 96-well plates were scanned at 8 and 24 h. Cellomics was employed to analyze the scanned images to assess the ability of cells to migrate.
9
Cell Migration Assays for Transfected Cells
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In order to analyze the migration ability of transfected cells in our research, wound-healing assay and transwell assay were performed. For wound-healing assay, lentivirus transfected A549 and NCI-H1299 cells (5 × 104 cells/well) were plated into 96-well plates for culturing. Scratches were made by a 96 wounding replicator (VP scientific). Photographs were taken by a fluorescence microscope at 0 h, 8 h, and 24 h and cell locations were recorded, respectively. Cell migration rates of each cell group were calculated. In transwell assay, cells were seeded into a 24-well plate in the upper chambers, and medium supplemented with 30% FBS was added into the lower chambers. Cells were fixed with 4% formaldehyde and stained by Giemsa and the migration ability of cells was analyzed.
10
Wound Healing Assay for Prostate Cancer
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Lentivirus-infected (shCtrl, shPSMC2) DU 145 and PC-3 cells (5 × 104 cells/well) were plated into a 96-well dish in triplicate for culturing. Scratches were made by a 96 wounding replicator (VP scientific, San Diego, CA, USA) across the cell layer with 90% confluence. Photographs were taken by a fluorescence microscope at 0 h, 24 h and 48 h after scratching. Cell migration rates of each group were calculated.
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