Is there any protocol or method to permeabilize suspension cell (k562 cell) without fixing them? I want to do live cell imaging hence I don't want any fixation?
If you really want to get proteins into live cells, you may have to look for options like protein-transfection or some specific approach. If you use any sort of detergent permeabilization you anyway will lose the cell integrity and will kill the cells
Electroporation wouldn't kill the cells. We also toyed with the idea of a gene gun at one point, but that is more for tissues perhaps.
Mild transfection reagents as you would use for transient transfections - fugene and the like. I did this years ago to get quantum dots into cells and tried various means. There are likely new and better products now.
Other ideas are Calcium phosphate...liposomes (lipofection).