Hi Malini, I've been studying hematological malignancies for years.

For suspension cells, firstly, you label cells with a fluorescent dye, such as PKH26, then you can use transwell system to evaluate the migrate capabilities by measuring the fluorescent intensity.

If you want to detect the migrated cells attached to the bottom, you can follow the same way. However, you need to pre-seed a monolayer of the BMSCs; after migration, you wash the suspension cells away softly in the bottom, then read the fluorescent intensity from the attached cells. This experiment is called cell adhesion assay, not invasion assay.

Actually, people are always confused about the definition of migration and invasion. For blood cells, I would like to use dissemination rather than invasion.

You can do the transwell experiment which can reflect cell migration. If you pre-seed BMSCs in the bottom, you can also detect cell adhesion.

Hope this information are helpful for you.

Jennifer

Hi Han Zhang , thank you very much for your reply. The use of the dye seems interesting and useful. May I ask, once the cells are labelled with the dye, how many passages can they undergo before losing the dye? Or must the cells be labelled and immediately be used for seeding? And when you say measure the fluorescence intensity, how do you do so in a 24-well format? Thank you very much for your help!

Hi Malini Rethnam , good questions! You can see the manual for PKH26 dye, which can, if my memory serves me right, last for weeks. As for the fluorescence intensity, you can use a fluorescent reader, which can read 6-, 12-, 24-, and 96-well plates. Before reading, you can choose which setting you'd like to read out, and just put the plate in it, that's it!

Hi Han Zhang , thank you very much for your assistance!