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
What are you trying to get into 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).
Low concentrations of saponin works well and their effects are reversible.
Nanotechnology. 2013 May 24;24(20):205101. doi: 10.1088/0957-4484/24/20/205101. Epub 2013 Apr 19.
A new technique for reversible permeabilization of live cells for intracellular delivery of quantum dots.
Medepalli K1, Alphenaar BW, Keynton RS, Sethu P.
Hi Badri, what do you want to "put inside"? If nanodots, they may go in without help.
Thanks everyone, actually i have a actin red dye which is not permeable to k562 cells and hence i would like to put this dye inside cells without killing them,ofcourse i know there are commercial available actin dyes which dont which are permeable BUT right now i would like use the one i have
do u think i have use mild conc of detergent like tween20 etc?
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