Does anyone know if there is a way to acquire slices for a Z-stack in random order?
Using multi-D acquisition always acquires slices from bottom (coverslip) to top of cell. I want to acquire those slices in random order.
Example: Acquire a through series z-stack of 100 different cells in a population. Stack for each cell would include 20 slices at 0.5 um/slice for 10 um total Z-axis distance. But each cell would have slices collected in random order. Is there a way to do this?
The reason is that I have a FRET pair comprised of fluorophores with different bleach rates. So when I acquire a Z-stack through multiple cells there is always an intensity bias for the first slice compared to the last slice. I thought if I collected slices in random order from 100 different cells the bias would disappear in the mean of all 100 Z-stacks.
Any suggestions would be welcome. Sorry did not fit well into any of the categories.