2.7. Spotting test
To compare the effect of manual and automatic counting, a spotting test was applied. Briefly, the concentration of yeast cells in the solution was calculated through the haemocytometer formula according to the results of the manual counting and the “Yeast Counter” macro. Samples with the maximum concentration were taken and diluted to 2×107 cells per ml, which were subsequently subjected to 10-fold dilutions. Five microlitres of each dilution, whose concentration ranged from 2×107 to 2×103 cells per ml, were spotted onto YPD solid media and incubated at 30 °C for 3 to 5 days. After incubation, comparisons between the manual and automatic groups were performed, in which extra care was taken for the spots with the fewest cells (Figure 2).