The 143 B and U2OS cells were seeded into 6-well plates overnight, then incubated with 150 μM GA for 48 h, and cell metastasis and cell invasion were detected. As mentioned before [20 (link)], the monolayer cells were carefully scratched by a sterile tip, and incubated with 150 μM GA in DMEM with 1% FBS. With 48 h-treatment, the wound areas were estimated by Image J. For transwell assays, GA-treated cells and control cells were seeded in the upper chamber with serum-free medium, and 10% FBS medium was added into the lower chamber. Then they were incubated for 24 h, and the cells on the upper chamber were gently scraped. The cells stayed on the lower membrane surface were fixed with methanol for 20min and stained with 0.1% crystal violet. Images were taken by Multifunctional Cell Imaging Microplate reader (BioTek, USA) and invasion rates were recorded by Gene 5.
Multifunctional cell imaging microplate reader
Manufactured by Agilent Technologies
Sourced in United States
The Multifunctional Cell Imaging Microplate Reader is a versatile lab equipment that combines cell imaging and microplate reading capabilities. It is designed to capture high-quality images and analyze various cellular parameters in a multi-well format.
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Lab products found in correlation
3 protocols using multifunctional cell imaging microplate reader
1
Evaluation of Ginkgolic Acid's Impact on Cell Metastasis and Invasion
2
Transwell Invasion Assay for Baicalein
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The transwell membranes were purchased from Corning Inc (New York, NY, USA). Transwell invasion assays were performed according to the manufacturer's protocol. Briefly, 1.0 × 104 baicalein-treated cells and control cells were resuspended in serum-free medium and seeded in the upper chamber, which was coated with Matrigel (Corning, USA). The low chambers were filled with the complete DMEM containing 10% FBS. After incubation for 48 h, cells on the upper surface were removed and the membrane were fixed with methanol for 20 min. Then the membrane was stained with 0.1% crystal violet, and images were captured by Multifunctional Cell Imaging Microplate reader (BioTek, USA). The invaded cells were counted in five different fields under microscopy.
3
Wound Healing and Transwell Assays
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For wound healing assays, cells were plated into 12-well plates and incubated overnight. To prevent the influence of cell proliferation, cells were pre-treated with mitomycin C (10 μg/ml) for 1 h, and then the monolayer was carefully scratched with a sterile plastic tip, and washed with PBS for three times to remove floating cells. Then the cells were cultured in the presence or absence of GA in serum-free media for 48 h, and images were captured under a microscope. The status of wound area was evaluated by ImageJ. The relative wound area was calculated according to the formula: S48/S0, where S0 and S48 are the wound area at 0 and 48 h respectively.
For transwell assays, 5.0 × 104 GA-treated cells and control cells were resuspended in serum-free medium and seeded in the upper chamber, which was coated with Matrigel (Corning, United States). Complete DMEM with 10% FBS was added into the lower chamber. After incubation for 20–36 h, the cells on the lower membrane were fixed with methanol for 20 min and stained with 0.1% crystal violet. Images were captured by Multifunctional Cell Imaging Microplate reader (BioTek, United States) and invaded cells were analyzed by using Gene5 software.
For transwell assays, 5.0 × 104 GA-treated cells and control cells were resuspended in serum-free medium and seeded in the upper chamber, which was coated with Matrigel (Corning, United States). Complete DMEM with 10% FBS was added into the lower chamber. After incubation for 20–36 h, the cells on the lower membrane were fixed with methanol for 20 min and stained with 0.1% crystal violet. Images were captured by Multifunctional Cell Imaging Microplate reader (BioTek, United States) and invaded cells were analyzed by using Gene5 software.
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