Cells were plated at 8,000 cells per well in a 96-well plate with 100 μl of medium per well, which were left overnight to equilibrate, and assessed for confluency (%) after 72 h following the drug treatments. Five images per 96 wells (×10 objective) were acquired using the IncuCyte® Live Imaging System (IncuCyte® Software (v2019B), Sartorius, Gottingen, Germany) and analyzed using the imaging analysis software. Six wells were used per treatment per experimental replicate.
Incucyte live imaging system
Manufactured by Sartorius
Sourced in United States
The IncuCyte live imaging system is a laboratory instrument that enables continuous, automated monitoring and analysis of cell cultures in an incubator environment. It provides real-time, quantitative data on various cellular processes and behaviors over an extended period of time.
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Lab products found in correlation
5 protocols using incucyte live imaging system
1
Cell Proliferation Assay with IncuCyte
2
PBMCs Activate Cytotoxicity Against Tumor
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Human PBMCs were isolated from the blood from healthy donors and activated with 100 ng/mL anti-CD3, 100 ng/mL anti-CD28, and 10 ng/mL IL2 (#317303; #302913; #589102) (BioLegend), and then co-cultured with tumor cells at 10:1 ratio. Cell death was assessed by a fluorescence caspase-3/7 substrate (#4440, Essen Bioscience) and monitored by the IncuCyte live imaging system (Sartorius).
3
SLURP1-Induced Morphological Changes
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SLURP1-induced morphological changes were evaluated using the IncuCyte live imaging system (Essen BioScience; Ann Arbor, MI, USA) in Caco2 cells cultured in 96-well plates at 1 × 104 cells/well. Cells were treated with rSLURP1 at 50, 200 nM and examined to investigate SLURP1-induced changes in cell morphology and cell proliferation for three days.
4
Scratch Wound Assay for Cell Migration
5
Invasion Assay for Brain Tumor Stem Cells
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Invasion assays were performed as previously described (Restall et al, J Vis Exp. 2018 Aug 29;(138)). In brief, Small BTSC spheres were collected form culture flasks and treated in 6-well plates. Spheres were treated with DMSO, 0.5 µM AMP001 or 0.25 µM Stattic for 24 hours.
Spheres were then transferred to 1.5 mL Eppendorf tubes to allow gravity pellets to form. Media was removed and spheres were resuspended in a Rat Collagen I (Cultrex) matrix. 100 μL/well of collagen suspended spheres was then transferred to a cold 96-well plate with 3 experimental replicates for each treatment condition. Plates were transferred to an Incucyte Live Imaging System (Essen Bioscience). Area of cellular invasion was recorded hourly for 24 hours.
Spheres were then transferred to 1.5 mL Eppendorf tubes to allow gravity pellets to form. Media was removed and spheres were resuspended in a Rat Collagen I (Cultrex) matrix. 100 μL/well of collagen suspended spheres was then transferred to a cold 96-well plate with 3 experimental replicates for each treatment condition. Plates were transferred to an Incucyte Live Imaging System (Essen Bioscience). Area of cellular invasion was recorded hourly for 24 hours.
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