Cell Biology Scratching Cos7 monolayer –
“Pushing” on cells with a coverslip – Mechanical forces
Cos7 H250 WT cells – Mechanical stimulation with MWG
Cos7 H250 GNAS KO + WT cells – Mechanical stimulation with MWG
cAMP and Calcium simulataneous FLIM imaging – Response of cell monolayer to scratch
External mechanical stimulator- imaging based pooled screen to stimulate entire dish of cells
Cell signaling dynamics – studied in real time at single cell level using FRET-FLIM
Image analysis – selection of cells that are “hits”
“Photoactivation” – using a 405nm laser to activate PA-mCherry in single cell nuclei
Cells imaged and segmented – All cells imaged for the mechano imaging based pooled screen Replicate1
Pipeline of an imaging based pooled CRISPR screen
