NF-κB moving into the nucleus is one of the most studied events in inflammation biology. YAP/TAZ shuttling is the readout of the Hippo pathway, critical in cancer and… See the analysis in action: https://youtube.com/watch?v=kDdKmLDTt1U
NF-κB moving into the nucleus is one of the most studied events in inflammation biology. YAP/TAZ shuttling is the readout of the Hippo pathway, critical in cancer and regeneration. FOXO translocation reports on insulin signaling. These are among the most common signaling pathway readouts in drug discovery, and they all come down to one number: the nuclear-to-cytoplasmic intensity ratio.
The technical challenge is defining where to measure the intensities. You first need to mask the nucleus (from DAPI), then a cytoplasmic mask, then intensity measurements in both regions. It's doable in general-purpose tools, but it requires configuration, scripting or pipeline building for every new experiment.
Cytely automates the mask generation: segment nuclei and cytoplasm, subtract the nuclei, measure target channel intensity in both compartments, compute the ratio. The result is a per-cell N/C ratio you can plot against dose to build translocation curves.
How the assay runs
Segment nuclei from DAPI → dilate to approximate cell boundary → subtract nuclei to create cytoplasmic ring → measure target channel intensity in both compartments → compute ratio.
What you provide
- Ch1: DAPI (nuclear stain)
- Ch2: target protein (IF or fluorescent tag)
What you get
- Nuclear-to-cytoplasmic intensity ratio (N/C ratio) per cell
- % translocated cells per condition
Ready to run
YAP, NF-κB, FOXO, p65, STAT3, any nuclear/cytoplasmic shuttling protein.
Inquire for support
Organelle-specific translocation (e.g., ER-to-Golgi).