Cells were seeded in a 6-well plate (6×104 cells/well) and were cultured for 24 h. The cells were then exposed to PBS (control), or were transfected with miRNA NC or miR-214 inhibitor. Subsequently, the cells were scratched using a 200-µl pipette tip (Thermo Fisher Scientific, Inc.). Following culturing at 37°C for 24 h, the cells were observed under a DSX100 optical microscope (Olympus Corporation, Tokyo, Japan).
Dsx100 optical microscope
Manufactured by Olympus
Sourced in Japan
The Olympus DSX100 is an optical microscope designed for routine observation and analysis. It features high-quality optics, simple operation, and a compact design. The DSX100 provides clear, high-contrast images for a variety of applications.
Automatically generated - may contain errors
Lab products found in correlation
Gfr matrigel, by BD (2 mentions)
Transwell chamber, by Corning (2 mentions)
Image pro plus 6.0 analysis software, by Media Cybernetics (1 mentions)
Tak1 antibody, by Merck Group (1 mentions)
Matrigel, by BD (1 mentions)
Transwell chambers with 8 m pore filters, by Corning (1 mentions)
8 m pore filter, by Corning (1 mentions)
Matrigel, by Corning (1 mentions)
Matrigel, by Merck Group (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
H9c2 rat embryo cardiomyocytes, by American Type Culture Collection (1 mentions)
Transwell chambers containing 8 m pore filters, by Corning (1 mentions)
10 protocols using dsx100 optical microscope
1
Wound Healing Assay with miR-214 Inhibitor
2
Immunohistochemical Analysis of TAK1 in Kidney
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The kidney tissue sections were deparaffinized and antigen retrieval was performed using a microwave oven at high temperature (300 W) for 30 min. Following the exhaustion of endogenous peroxidase with methanol and hydrogen peroxide for 30 min at room temperature, the slides were blocked with 0.5% bovine serum albumin (Sigma-Aldrich; Merck KGaA) for 60 min at 37°C and incubated with TAK1 antibody (1:200 dilution; cat. no. ab109526; Abcam) overnight at 4°C. The SABC staining kit (cat. no. SA1026; Boster Biological Technology) was used to perform the chromogenic reaction. Following rinsing in PBS, the samples were incubated with the mouse anti-rabbit antibody (cat. no. SA1026; Boster Biological Technology) at 37°C for 30 min, followed by rinsing in PBS three times for 5 min. The samples were covered in a drop of DAB developing solution, cell nuclei were stained with hematoxylin at room temperature for 5 min, following dehydration with an alcohol gradient, the samples were cleared with dimethylbenzene xylene and mounted with neutral gum sealant. Photomicrographs of different fields of view were captured using an Olympus DSX100 optical microscope (Olympus Corporation; magnification, ×400). and the average optical density (OD) was calculated using Image-pro plus 6.0 analysis software (Media Cybernetics, Inc.).
3
Transwell Invasion Assay for Cells
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Matrigel (200 µg/ml, 100 µl; BD Biosciences, Franklin Lakes, NJ, USA) was added to the upper Transwell chamber at room temperature until it solidified, after which, cells were added to the upper chamber. The medium supplemented with 15% FBS was added to the lower chamber. The transfected cells were centrifuged at 5,000 × g for 20 min at 4°C, and the cell suspension (2×105 cells/ml) was cultured in the upper chamber at 37°C for 24 h. Crystal violet (0.1%; Shanghai Gefan Biotechnology Co., Ltd., Shanghai, China) was used to stain the cells for 20 min. Finally, cells were observed under a DSX100 optical microscope (Olympus Corporation).
4
Wound Healing Assay for Cell Metastasis
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Wound healing assay was applied to determine cell metastatic abilities. To be more specific, 1×105 SKOV3 cells were treated with different concentrations of ginsenoside Rg3 (0, 50, 100 and 200 µg/ml). Next, the cells were inoculated in each well of 12-well plates and scratched gently to form a cell-free area and then cultured in an incubator for 12 and 24 h at 37°C. Finally, the diameters of cell-free areas were assessed under an Olympus DSX100 optical microscope (Olympus Corp., Tokyo, Japan).
5
Wound Healing Assay for Cell Metastasis
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Wound healing assay was used to determine cell metastasis abilities. To be more specific, 1×105 cells were inoculated in each well of 12-well plates and incubated at 37°C for 24 h. Next, the confluent monolayer cells were first scratched gently to form a cell-free area and then cultured in an incubator at 37°C for 24 h. Finally, the diameters of cell-free areas were measured under Olympus DSX100 optical microscope (Olympus Corporation, Tokyo, Japan).
6
Measuring VSMC Invasion Capacity
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The invasive abilities of HA-VSMCs were measured using 24-well Transwell® chambers with 8-µm pore filters (Corning Inc). Cntl, EV and si-COL6A1 cells (5×104 cells) treated or untreated with 20 ng/ml PDGF-BB, diluted in Medium 231 supplemented with SMGS, were seeded into the Matrigel® GFR (BD Biosciences)-coated Transwell® upper chambers, with the coating process at 37°C for 30 min. The lower chambers were filled with Medium 231 supplemented with SMGS. Following 24 h incubation, the Transwell membranes were stained using 0.1% crystal violet for 30 min at 37°C. The number of invasive cells in random 5 fields was then calculated from images captured using Olympus DSX100 optical microscope (Olympus Corporation; magnification, ×200).
7
Ginsenoside Rg3 Inhibits Ovarian Cancer Cell Invasion
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Cell invasion abilities of ovarian cancer cells treated with ginsenoside Rg3 were assessed using 24-well Transwell chambers that contained 8-µm pore filters (Corning Inc., Corning, NY, USA). In brief, 5×104 SKOV3 cells treated with different concentrations of ginsenoside Rg3 (0, 50, 100 and 200 µg/ml) were cultured in DMEM culture media in Matrigel GFR-coated (BD Biosciences) upper chambers of the Transwell. In addition, DMEM culture media containing 10% FBS was filled into the lower chambers. After having been incubated for 48 h, the bottom membrane was stained with 0.1% crystal violet for 30 min at 37°C. The number of invasive cells was calculated using Olympus DSX100 optical microscope (Olympus Corp.) with a magnification of ×200.
8
Matrigel-Transwell Cell Invasion and Wound Healing Assays
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Matrigel (100 µl; Sigma-Aldrich; Merck KGaA) was diluted in free-FBS RPMI-1640 medium at a ratio of 1:8 and added to the Transwell chamber (Corning Incorporated) in an incubator for 6 h at 37°C to ensure solidification of the Matrigel. The cells (1×106/ml) were resuspended with free-FBS RPMI-1640 medium and were then added in top chamber of the Transwell following treatment with the reagents for 48 h, and 20% FBS RPMI-1640 medium was added to lower chamber of the Transwell. The Transwell containing the cells was placed in an incubator at 37°C for 24 h. A methanol:acetone (1:1) solution (Beijing Solarbio Science & Technology Co., Ltd., Beijing, China) was used to fix the cells, and 4% crystal violet solution (Beijing Solarbio Science & Technology Co., Ltd.) was used to stain the invaded cells. Images were captured using Olympus DSX100 optical microscope (Olympus Corporation, Tokyo, Japan).
The reagents were used to treat the cells for 48 h, following which the cells were resuspended in 2% FBS culture medium. The cells were added to a 35-mm dish, and a 200 µl pipette tip (Sigma-Aldrich; Merck KGaA) was used to scratch the cells. After 48 h, images of the scratches were captured using a microscope. The shortened distances of the scratches were used to determine the migration rates.
The reagents were used to treat the cells for 48 h, following which the cells were resuspended in 2% FBS culture medium. The cells were added to a 35-mm dish, and a 200 µl pipette tip (Sigma-Aldrich; Merck KGaA) was used to scratch the cells. After 48 h, images of the scratches were captured using a microscope. The shortened distances of the scratches were used to determine the migration rates.
9
H9c2 Cardiomyocyte Culture Protocol
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H9c2 rat embryo cardiomyocytes purchased from the American Type Culture Collection (Manassas, VA, USA) were cultured in Dulbecco's modified Eagle's medium (DMEM; Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) supplemented with 10% fetal bovine serum (Gibco; Thermo Fisher Scientific, Inc.) and 100 U/ml penicillin/streptomycin (Invitrogen; Thermo Fisher Scientific, Inc.) at 37°C with 5% CO2. Cells at logarithmic phase were used in the current study. H9c2 cell morphology was identified under an Olympus DSX100 optical microscope (Olympus Corporation, Tokyo, Japan) at 48 h following treatment (magnification, ×100 or 200).
10
Transwell-Based Cell Invasion Assay
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Using 24-well Transwell chambers containing 8-µm pore filters (Corning Incorporated, Corning, NY, USA), cell invasion abilities of cervical cancer cells with KLF1 interference were compared to Control and Mock groups. To explain, 5×104 cells were cultured in Matrigel GFR (BD Biosciences, Franklin Lakes, NJ, USA)-coated Transwell upper chambers using DMEM culture media. Meanwhile, DMEM culture media containing 10% FBS was filled in the lower chambers. After the incubation at 37°C for 24 h, the bottom membrane was stained with 0.1% crystal violet at 37°C for 30 min. Cell numbers was calculated using Olympus DSX100 optical microscope (Olympus Corporation) with the magnification set at 100-fold.
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