Human SMCs and ECs were plated separately in a 96-well format and grown overnight. Upon reaching confluency cells were subjected to automated scratch assay using Incucyte plate scratcher. The plates were subsequently was imaged using an Zoom (Essen Bioscience, Ann Arbor, MI) every 2 h for 30 hours. Cells were imaged under phase contrast as well as fluorescence. Analysis was performed using Incucyte’s Imaging software. The IncuCyte ZOOM Scratch Wound assay utilizes the WoundMaker-IncuCyte ZOOM-ImageLock Plate system to analyze both 2D-migration and 3D-invasion in label-free, live cells (Essen Bioscience ). Wound Confluence is a report of the confluence of cells within the wound region, given as the percentage of the wound region area occupied by cells. Relative Wound Density(RWD) relies on measuring the spatial cell density in the wound area relative to the spatial cell density outside of the wound area at every time point. The metric is self-normalizing for changes in cell density which may occur outside the wound due to cell proliferation and/or pharmacological effects. Importantly, the RWD metric is robust across multiple cell types as it does not rely on finding cell boundaries.
Woundmaker incucyte zoom imagelock plate system
Manufactured by Sartorius
Sourced in United States
The WoundMaker-IncuCyte ZOOM-ImageLock Plate system is a lab equipment product that combines two key components: the WoundMaker and the IncuCyte ZOOM with ImageLock Plate. The WoundMaker is a tool used to create standardized, reproducible wounds or barriers in cell monolayers. The IncuCyte ZOOM is an automated, live-cell imaging system that allows for continuous monitoring and analysis of cell migration and wound healing over time. The ImageLock Plate is a specialized multi-well plate designed to work with the IncuCyte ZOOM, enabling the capture of high-quality, time-lapse images of the cell monolayer and wound area.
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Lab products found in correlation
2 protocols using woundmaker incucyte zoom imagelock plate system
1
Automated Scratch Assay for Cell Migration
2
In Vitro Scratch Wound Assay for Evaluating SEA Effects
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In vitro scratch wound assays were carried out to determine the effect of SEA extracted from control and CRISPR/Cas9 mutated eggs [80, (link)81] (link). HSC cells (LX-2) were cultured in completed medium containing high glucose EMDM (Invitrogen), 2% fetal bovine serum (FBS), 1% glutamax and 1% Pen/strep. Cells were grown to create a confluent monolayer in 96-well plates.
Then the monolayers were scraped in a straight line to create a "scratch" using the Wound Maker-IncuCyte ZOOM-Image Lock Plate system (Essen Bioscience, Michigan, USA) and washed once with the culture medium. Cells were continued to culture in the fresh completed medium containing 30 µg/ ml SEA. Residual endotoxin was assessed by using an Endotoxin Standards kit (Lonza, Anaheim, USA) to ensure there was no LPS contamination in the SEA extracted from control and AChE-KI eggs). The growth of cells was monitored by IncuCyte Zoom for 3 days. Images were acquired for each well at 3 hours interval by an in-built phase contrast microscope. The area of the scratch was measured using ImageJ (image processing program, Java). The total area (μm 2 ) was obtained at every time point until would closure, and triplicate measures were compared to calculate an average closure rate per group (area of original wound/time to closure in hours). All data were then statistically analysed using GraphPad Prism Software V7.
Then the monolayers were scraped in a straight line to create a "scratch" using the Wound Maker-IncuCyte ZOOM-Image Lock Plate system (Essen Bioscience, Michigan, USA) and washed once with the culture medium. Cells were continued to culture in the fresh completed medium containing 30 µg/ ml SEA. Residual endotoxin was assessed by using an Endotoxin Standards kit (Lonza, Anaheim, USA) to ensure there was no LPS contamination in the SEA extracted from control and AChE-KI eggs). The growth of cells was monitored by IncuCyte Zoom for 3 days. Images were acquired for each well at 3 hours interval by an in-built phase contrast microscope. The area of the scratch was measured using ImageJ (image processing program, Java). The total area (μm 2 ) was obtained at every time point until would closure, and triplicate measures were compared to calculate an average closure rate per group (area of original wound/time to closure in hours). All data were then statistically analysed using GraphPad Prism Software V7.
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