Severe cell detachment around the scratch

Symptom

After the scratch, the area around the line is littered with detached/floating cells, making the field of view dirty and obscuring later observations.

1. 1 Scratch was made with excessive force
2. 2 PBS wash was too aggressive
3. 3 Cells adhere poorly or are in suboptimal condition

1. 1 Scratch gently and avoid repeated scraping over the same line
2. 2 Pipette wash buffers gently against the well wall, not directly onto the monolayer
3. 3 Use cells in good adherent condition (passage number and confluence matter)

Related Video (2)

YouTube (Curated Tutorials) ★ 88

Scratch Assay Tutorial

"Direct scratch assay tutorial demonstrating the technique, most likely to show correct scratching method and avoid excessive force."

YouTube (Curated Tutorials) ★ 80

Wound healing

"Wound healing assay setup and analysis video covering how to perform the technique where this detachment failure occurs."