Hopefully, this will help you calculation inoculum dilutions easily.

Column A is self-explanatory
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Columns B and C are for spore counts taken from a hemacytometer o The count should cover the entire central grid (refer to “Hemacytometer”)

Column D calculates the concentration of your inoculum solution o =(SUM(B2:C2))/2/0.1
 This formula calculates the average of both of you hemacytometer counts and divides it by the volume of the central grid (0.1 µ l)

Column E is for germination percentage, calculated following the protocol “Germination and
bias assays” o If you assumed 100% germination, fill in “1” for all samples

Column F calculates the amount of triton water you must add to dilute to exactly 500 spores/ µ l o 500 spores/ µ l is more than enough for inoculating a seedling o =(D2*182/(500/E2))-182
 The numerator in parentheses calculates the number of spores in your solution.
It multiplies concentration by 182 because presumably there were 200 µ l of
Triton water to being in your eppendorf tube if you follow “Single-meristem
stage inoculation,” and you performed two counts which require 9 µ l each.
 The denominator calculates the final volume necessary to reach 500 spores/ µ l, corrected for germination percentage
 The subtraction accounts for current tube volume.

MAKE SURE THAT YOUR FINAL VOLUME OF INOCULUM IS ENOUGH TO INOCULATE ALL OF
YOUR SEEDLINGS WITH 2 µ l EACH