Comparison of Counting Standard Microsphere Dispersion and Concentration for enumerating Saccharomyces cerevisiae
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Abstract:
The concentration and statistical numbers of counting standard microspheres used for cell counting of the Saccharomyces cerevisiae strain (ATCC9080) were selected for analyzing the effects of microsphere aggregation. The dispersion, concentration, and statistical numbers of microspheres in Tween 20 or Tween 80 suspension for S. cerevisiae counting were evaluated. In addition, we determined the linearity between standard microsphere counting and plate counting. Our study showed that the dispersion of microspheres in 0.08% Tween 20 or 0.06% Tween 80 suspension was suitable, with a dispersion index of 3.30. For counting 105, 106, and 107 cells/mL of S. cerevisiae, the total statistical numbers of standard microspheres should not be less than 2 000, 1 500, and 1 000, respectively. In addition, the total statistical numbers of cells should not be fewer than 200, 400, and 2 600. At 105 to 107 cells/mL of S. cerevisiae, the counting results by standard microspheres were consistent with those of the plate counting (R2=0.996 9), with 106 beads/mL of standard microsphere being the most suitable concentration. Our results indicated the standard microspheres could be effectively dispersed by Tween 20 or Tween 80. Hence, microbial cells in suspension can be accurately counted by using an appropriate concentration of dispersed standard microspheres. The suitable parameters of the standard microspheres for microbial counting were identified to provide the application conditions of standard microspheres in GB/T 39730-2020.
