Accuracy here refers to how close is the estimation to the actual number of cells.
Basically, you have to count (estimate) how many target cells are present within an area. The most direct way is obviously counting all of them. This is impractical because the field of view of an instrument is limited in scope. Ignoring other complications, doing so would involve counting hundreds of thousand of FoVs. No one has time for that.
Alternatively, you can assume that any random FoV contains approximately the same number of target cells and multiply up to find the total. Of course, this assumption is wrong because density is not uniform. BUT if you have enough samples (n>>>1), the estimation becomes more accurate. That is the essence of the question. Currently, n = 30.
A: Not really. There is a limit to how much you can magnify. Like I said, you need hundreds of thousand of FoVs to cover everything. So unless you can magnify to the point where the covered areas are like orders of magnitude as large as the original, it makes no difference.
B: Yes, if n> 30, the average will be more representative.
C: Doing this will exacerbate the error.
D: There is no indication that there is a difficulty in identifying the target cells. At most, this will increase precision, not accuracy.
Edit: I never did this experiment but when I count, my n is like.... 4. N = 30 is definitely overkill 😀
