To perform the cell migration assay, cell monolayers with or without drug treatment were incubated in 24-well tissue culture plates in serum-free media for 45 minutes. In the experiment, a 1-mm wide model wound was created by scratching the cell monolayer with a sterilized 1000 μL pipette tip. In the leader cell formation experiment, cells were refreshed with DMEM containing serum. Time-lapse microscopy with bright-field and fluorescence imaging was performed using a microscope stage-top incubator from Okolab (NA, Italy). For migration assays involving drug treatments, cells were maintained with the same concentration of drugs throughout the experiment, and the migration of the epithelia was monitored after performing the scratch assay.
Microscope stage top incubator
Manufactured by Okolab
Sourced in Italy
The Microscope Stage Top Incubator is a device designed to maintain a controlled environment for samples under a microscope. It provides precise temperature regulation to ensure optimal conditions for live cell imaging and analysis.
Automatically generated - may contain errors
Lab products found in correlation
Dmi8 automated platform microscope, by Leica (4 mentions)
Growth factor reduced matrigel, by Corning (4 mentions)
Las x software, by Leica (4 mentions)
Sulforaphane sfph, by Merck Group (3 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions)
Egm 2, by PromoCell (1 mentions)
6 protocols using microscope stage top incubator
1
Cell Migration Assay and Wound Healing
2
Cell Migration Assay and Wound Healing
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To perform the cell migration assay, cell monolayers with or without drug treatment were incubated in 24-well tissue culture plates in serum-free media for 45 minutes. In the experiment, a 1-mm wide model wound was created by scratching the cell monolayer with a sterilized 1000 μL pipette tip. In the leader cell formation experiment, cells were refreshed with DMEM containing serum. Time-lapse microscopy with bright-field and fluorescence imaging was performed using a microscope stage-top incubator from Okolab (NA, Italy). For migration assays involving drug treatments, cells were maintained with the same concentration of drugs throughout the experiment, and the migration of the epithelia was monitored after performing the scratch assay.
3
Angiogenesis Assay with Ov-GRN-1
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Growth factor-reduced Matrigel (Corning, Corning, NY, USA) was plated into a 96-well μ-angiogenesis plate (ibidi, Planegg, Germany) at 10 μl/well, and incubated at 37°C in 5% CO2 in air for 60 min as described (14 (link)). HUVECs were detached (above) and resuspended in complete endothelial cell growth medium 2 (EGM-2) (PromoCell), and seeded at 10,000 cells/well in medium supplemented with 10 µM sulforaphane (SFPH, Sigma) (negative control), 1.2 nM VEGF-165 (Novus Biologicals) (positive control), or 5, 10, 20, and 40 nM Ov-GRN-1. The ibidi plate was incubated for 12 h in a humidified atmosphere of 5% CO2 in air at 37°C in a microscope stage top incubator (OKOLAB, Pozzuoli, Naples, Italy). At intervals, photomicrographs of cells and nascent and developed tubules were collected using a Leica DMi8 automated platform microscope under bright field at 2.5 × magnification, and Leica LASX software (Leica).
4
Angiogenesis Assay with Sulforaphane
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Growth factor-reduced Matrigel (Corning, Corning, NY, USA) was plated into a 96-well μ-angiogenesis plate (ibidi, Planegg, Germany) at 10 μl/well, and incubated at 37 °C in 5% CO2 in air for 60 min as described [18 ]. 3B-11 cells were detached using Trypsin/EDTA and resuspended in DMEM (Gibco), and seeded at 30,000 cells/well in medium supplemented with 10 μM sulforaphane (SFPH, Sigma) (negative control), or 1.8 μg/mL or 3.6 μg/mL IPSE. The ibidi plate was incubated for 5 h in a humidified atmosphere of 5% CO2 in air at 37 °C in a microscope stage top incubator (OKOLAB, Pozzuoli, Naples, Italy). At intervals, photomicrographs of cells and nascent and developed tubules were collected using a Leica DMi8 automated platform microscope under bright field at 2.5 × magnification, and LASX software (Leica).
5
In Vitro Angiogenesis Assay with Sulforaphane and IPSE
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Growth factor-reduced Matrigel (Corning, Corning, NY, USA) was plated into a 96-well µ-angiogenesis plate (ibidi, Planegg, Germany) at 10 µl/well, and incubated at 37 °C in 5% CO 2 in air for 60 min as described (18). 3B-11 cells were detached using Trypsin/EDTA and resuspended in DMEM (Gibco), and seeded at 30,000 cells/well in medium supplemented with 10 µM sulforaphane (SFPH, Sigma) (negative control), or 1.8 µg/mL or 3.6 µg/mL IPSE. The ibidi plate was incubated for 5 h in a humidi ed atmosphere of 5% CO 2 in air at 37 °C in a microscope stage top incubator (OKOLAB, Pozzuoli, Naples, Italy). At intervals, photomicrographs of cells and nascent and developed tubules were collected using a Leica DMi8 automated platform microscope under bright eld at 2.5 × magni cation, and LASX software (Leica).
6
Matrigel-based In Vitro Angiogenesis Assay
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Growth factor-reduced Matrigel (Corning, Corning, NY, USA) was plated into a 96-well μ-angiogenesis plate (ibidi, Planegg, Germany) at 10 μl/well, and incubated at 37°C in 5% CO 2 in air for 60 min as described (18). 3B-11 cells were detached using Trypsin/EDTA and resuspended in DMEM (Gibco), and seeded at 30,000 cells/well in medium supplemented with 10 µM sulforaphane (SFPH, Sigma) (negative control), or 1.8 μg/mL or 3.6 μg/mL IPSE. The ibidi plate was incubated for 5 h in a humidi ed atmosphere of 5% CO 2 in air at 37°C in a microscope stage top incubator (OKOLAB, Pozzuoli, Naples, Italy). At intervals, photomicrographs of cells and nascent and developed tubules were collected using a Leica DMi8 automated platform microscope under bright eld at 2.5 × magni cation, and LASX software (Leica).
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