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Count the mean number of typical fluorescent cells per field of view (c).
Calculate the number of typical fluorescent cells per microscope slide window (C).
C = c × S/s
where | S | = | surface area of window of multispot slide |
and | s | = | surface area of objective field |
s = πi2/4G2K2 | where | i | = | field coefficient (varies from 8 to 24 depending upon ocular type) |
K | = | tube coefficient (1 or 1,25) | ||
G | = | magnification of objective (100 ×, 40 × etc.). |
Calculate the number of typical fluorescent cells per ml of re-suspended pellet (N).
N = C × 1 000/y × F
where | y | = | volume of re-suspended pellet on each window |
and | F | = | dilution factor of re-suspended pellet.” |