PE and AER

FACS is a fucking pain in the ass. I am not referring to the infect-your-cells-with-something-green-then-sort-out-the-green-ones kind of FACS - that's pretty easy. I am referring to staining cells with fluorescent-tagged antibodies, and then generating complicated gates based upon size and positive or negative signal from umpteen different antibodies. All of this staining and sorting is preceded by a variety of treatments to the whole cell population that take a non-trivial amount of time. I should also mention this is all being done in a 96-well format, and the reading of said plate (equivalent to one clone's worth of cells) takes about two hours. I should also mention that there is always demand for an increasing number of clones to be run on any given day.

I have never, ever done an experiment where the perceived effort (let's call it PE) to accomplish the task is so tiny, where the actual effort required (AER) is so massive. Let me reiterate:

I have done chromatin IPs, mouse colony husbandry, immunocytochemistry, Southerns, qPCR, and nothing - nothing! - comes close to FACS in this equation above. Many experiments have a similar AER (chIP comes to mind), but the PE for those experiments is also much higher (e.g., "Oh, it would be nice if we had chIP data on X, but I don't know if you can get that much work done before [date needed]!"). Mouse colony husbandry (tagging, tailing, genotyping) is probably the closest (e.g., "Why are you spending so much time in the mouse room? How long can it take to tag all those mice?") but I'd say it's only two less-than signs.

Is there anything more horrible than FACS out there? ::shudder::