What does a wound healing assay tell you?

The wound healing assay is a standard in vitro technique for probing collective cell migration1-6 in two dimensions. In this assay, a cell-free area is created in a confluent monolayer by physical exclusion or by removing the cells from the area through mechanical, thermal or chemical damage.

What is cell migration in wound healing?

In essential physiological processes including morphogenesis, wound healing and tissue regeneration, cells often move as a tightly or loosely associated cohesive group. This type of migration, during which cells are influenced by the interaction with their neighbors, is known as ‘collective cell migration’.

How does a scratch assay work?

The scratch-wound assay is a simple, reproducible assay commonly used to measure basic cell migration parameters such as speed, persistence, and polarity. Cells are grown to confluence and a thin “wound” introduced by scratching with a pipette tip. Cells at the wound edge polarise and migrate into the wound space.

How do you do a scratch wound assay?

Scratch or wound assay

1. i. Scrape the cell monolayer in a straight line to create a “scratch” with a p200 pipet tip.
2. ii. To obtain the same field during the image acquisition, create markings to be used as reference points close to the scratch.
3. iii. Place the dish in a tissue culture incubator at 37 °C for 8–18 h.
4. iv.

What does colony formation assay mean?

Clonogenic assay
Clonogenic assay or colony formation assay (CFA) is an in vitro cell survival assay based on the ability of single cells to grow into colonies [1]. It is the gold standard to determine cell reproductive death after treatment with ionizing radiation.

What is cell migration assay?

The Cell Migration Assay measures the number of cells traversing a porous membrane, while the Cell Invasion Assays monitor cell movement through extracellular matrices.

How is wound healing assay calculated?

Wound healing assay Where Wi is the average of the initial wound width, Wf is the average of the final wound width both in μm and t is the time span of the assay in hours. Additionally, At = 0 is the initial wound area, At = Δt is the wound area after n hours of the initial scratch, both in μm2.

What is invasion assay?

The Cell Invasion Assays offer a flexible, standardized, high-throughput format for quantitating the degree to which invasive cells penetrate a barrier consisting of basement membrane components in response to chemoattractants and/or inhibiting compounds. …

What is the mechanism of MTT assay?

MTT (3-(4,5-DIMETHYLTHIAZOL-2-YL)-2,5-DIPHENYLTETRAZOLIUM BROMIDE) ASSAY. The MTT assay relies on the cellular reduction of tetrazolium salts to their formazan crystals. The range of cell concentrations in which there is a direct relationship between optical density and the number of cells counted should be determined.

How MTT assay is performed?

MTT Assay Protocol

1. Prepare cells and test compounds in 96-well plates containing a final volume of 100 µl/well.
2. Incubate for desired period of exposure.
3. Add 10 µl MTT Solution per well to achieve a final concentration of 0.45 mg/ml.
4. Incubate 1 to 4 hours at 37°C.