3 replies to this topic

[science noob]

I understand that flow cytometry is a quantitative method to look at protein expression but would suspended cells (spherical) share similarities seen in monolayer attached cells?

Would trypsinising cells for the purposes of immuno-positivity staining in flow disrupt cellular structure integrity?

[Rsm]

FACS is mostly intended to analyze expression of surface markers in suspended cells (ie hematopoietic cells). It is rather challenging to detect intracellular proteins, if they are not labelled by GFP. Trypsinizing always leads to a change in surface molecule composition, otherwise you couldn't detach the cells. But the intracellular structure should not be affected.

[Matrixwoo]

flow cytometry is the technique for counting or examining cells and chromosomes by suspending them in a stream of fluid and passing them by an electronic detection apparatus.
immunostaining  applies to any use of an anti-body method to detect a specific protein in a sample

[science noob]

In a way, flow cytometry is more quantitative vs. conventional staining. But some say that flow is an immunologists' pet 'toy' whilst slide-based immunostaining and Western blot are "preferred" by cell biologists and biochemists. Any thoughts?