Best method to measure force transmission between cells
Posted by Joana Carvalho, on 6 April 2015
We have been trying to implement FRET in the lab and transfect our cells with mechanical biosensors, but so far we haven’t managed to successfully force our cells to express our constructs. After some digging, we realised that this technique does not work for all cell types. Its success seems to be highly dependent on cell sensitivity, which may be one of the reasons why we are not getting positive results out of it because HUVECS are pretty sensitive cells to work with.
That’s why I am here asking for advice regarding alternative techniques we could use to measure cell-cell force transmission. Fom what I have seen so far, a lot of people have been using AFM (Atomic Force Microscopy) to quantify cell adhesion to the ECM. However, what I am interested in is actually to measure force between neighbour cells. There are also other techniques to consider like micropost arrays or even flow chamber assays, but these methods kind of escape what we would like to address. Anyone has any other advice for something else we could try?
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Try TFM. Work by Margaret Gardel.
Thank you for the recommendation! I’ll definitely take a closer look at her work on TFM.
If your cells can be suspended, a dual pipette assay may be best.
You can measure the absolute value of the force holding cells together independently of any adhesion to a substrate while imaging your sample.
Check out my protocol:
Dual pipette aspiration: a unique tool for studying intercellular adhesion Biro M and Maître JL
Method Cell Biol (Protocol) 2015
One of its main application:
Adhesion functions in cell sorting by mechanically coupling the cortices of adhering cells
Maître JL, Berthoumieux H, Krens SFG, Salbreux G, Jülicher F, Paluch E, Heisenberg CP
Science. 2012
And a review of different methods to measure force transmission between cells
The role of adhesion energy in controlling cell-cell contacts
Maître JL, Heisenberg CP
Curr. Opin. Cell Biol. 2011
We are working with HUVECS that are adhesive cells, but according to my boss they should be able to form AJs while in suspension, so that shouldn’t be an issue.
I guess the biggest challenge for us would be establishing the experimental setup itself. I still haven’t had time to read your protocol in detail, but I already noticed it is fairly complex.
Would you be available for helping us with some technical details or doubts we may have if we decide to give it a try?
Thank you so much for all the help!