In wound-healing assay, 7.5X104 cells were seeded in Culture-Insert 2 well in μ-Dish 35 mm (Ibidi) and allowed to form monolayer. The insert was then removed and the migration area in the wound was imaged and evaluated in an IncuCyte S3 Live-Cell Analysis System (Sartorius) constantly for up to 3 days. Invasion assay was performed as previously published (16 (link)). Cells (2-2.5X104 for KPL4 and 3.5X104 for FC-IBC02) were plated in 6.5 mm Transwell champers (Sigma). 6 to 18 h later, depending on the invasive potential of the cell line, the cells that had invaded through the filter and attached to its bottom surface were stained with crystal-violet, imaged and counted in five independent fields/well using a Nikon Eclipse TE2000-U microscope equipped with a DS-FI3 Camera and NIS-Element Software (Nikon).
Protocol detail
Monitoring Cell Migration and Invasion Dynamics
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Corresponding Organization : University of Pennsylvania
Other organizations : Cedars-Sinai Medical Center, The University of Texas MD Anderson Cancer Center
Top 5 similar protocols
Variable analysis
independent variables
- Cell seeding density (7.5 x 10^4 cells)
- Culture-Insert 2 well in μ-Dish 35 mm (Ibidi) used for wound-healing assay
- Transwell chambers (Sigma) used for invasion assay
- Cell lines (KPL4 and FC-IBC02) used for invasion assay
- Migration area in the wound (measured in wound-healing assay)
- Number of cells that invaded through the filter and attached to its bottom surface (measured in invasion assay)
- Monolayer formation prior to wound-healing assay
- Time period for wound-healing assay (up to 3 days)
- Time period for invasion assay (6 to 18 hours, depending on cell line)
- Staining of invaded cells with crystal-violet in invasion assay
- Imaging and counting of invaded cells using Nikon Eclipse TE2000-U microscope and NIS-Element Software
- Positive control: None specified
- Negative control: None specified
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
Sign up for free or login to display all annotations
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!