Severe cell detachment around the scratch (Wound Healing Assay)

Question

Accepted Answer

After the scratch, the area around the line is littered with detached/floating cells, making the field of view dirty and obscuring later observations. Common causes: 1) Scratch was made with excessive force 2) PBS wash was too aggressive 3) Cells adhere poorly or are in suboptimal condition Solutions: 1) Scratch gently and avoid repeated scraping over the same line 2) Pipette wash buffers gently against the well wall, not directly onto the monolayer 3) Use cells in good adherent condition (passage number and confluence matter)