I want to separate dead cells from a bacterial population after treatment of any drug or antimicrobial agent. Which technique will be more efficient FACS or MACS if I use PI and SYTO9 dye ? Is there any other technique to separate dead cells from live cells?
Hello Rakesh,
If you want to separate a population using fluorescent dyes, you must do cell sorting (FACS). Otherwise, you need a magnetic separation kit that allow you to specifically bind dead cells (it's negative selection) through beads-bounded antibodies (MACS).
Cheers,
Yamile
Hello Rakesh Mohapatra
The gold standard for cell separation is FACS. It is capable of processing millions of cells and isolating multiple, high purity subpopulations.
On the other hand, the related technology, (MACS), relies on direct (primary antibody-conjugated microbead) or indirect (primary antibody plus secondary antibody-conjugated microbead) magnetic labeling of cells prior to separation in a magnetic field. MACS is also used to select for cell populations using surface markers. However, it lacks the sensitivity and cell-specific data provided by a fluorescence-based system and is not easily compatible with multiple-marker profiles.
Moreover, MACS is often time-consuming and laborious. The hidden costs can be significant such as separation magnets, magnetic beads, and column replacements can all add up to the cost. MACS requires researchers to use separate columns and magnets for each experiment. The most significant disadvantage, though, is the effect magnetic fields have on some cell populations. For delicate cells, magnetic beads can cause damage.
I suggest you go for FACS since you already have the fluorescent dyes.
There are other techniques to separate dead cells from live cells, one of which I have come across is attached below for your reference.
Article Dielectrophoretic Concentration and Separation of Live and D...
Best.
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