I was washing cells with 1500 rpm (= 210 x g) after antibody staining for cytometric analysis. I didn't notice that one of my colleague changed the centrifuge machine setting to 6000 x g. I'm wondering:
1. The cells were previously fixed and permeabilized for intracellular staining, but after staining I didn't fix them (was gonna fix them again after two washes.) Is it possible the antibodies be dissociated from the cells/antigens simply by a centrifugation force of 6000 x g?
2. Although the cells were fixed, is 6000 x g large enough to destruct cell structure?
3. I was taught by my advisor that fluorochromes are easily dissociated from antibodies so always put them on ice and try not to frequently freeze and thaw. With a force of 6000 x g, will the fluorescent dyes detach from the antibody?
I'm gonna analyze these cells on a cytometer next week; just curious to know if antigen/antibody/fluorescence can be dissociated so easily. Thank you all for reading/answering this.