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Mixing Is Dispensable for Optical Density-Based High-Throughput Growth Screening Assay in Fission Yeast.
Kim Kiat Lim1, Jiunn Jye Chung2, Sha Ma2
1Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117596, Singapore.
International Journal of Molecular Sciences
|May 4, 2026
Summary
Mixing yeast cells before optical density measurement is not essential for high-throughput screening (HTS). Our study found no significant difference in cell growth dynamics, simplifying HTS workflows and improving efficiency.
Area of Science:
- Microbiology
- Biotechnology
- Pharmaceutical Sciences
Background:
- Optical density (OD) measurements are standard for assessing cell growth in high-throughput screening (HTS) for drug discovery.
- Resuspending cells before OD measurement is a common practice, but its necessity in HTS has not been rigorously evaluated.
Purpose of the Study:
- To systematically investigate the impact of cell mixing on OD-based growth measurements in yeast.
- To determine if omitting the mixing step affects the biological interpretation of HTS data.
Main Methods:
- Growth dynamics of *Schizosaccharomyces pombe* strains were measured with and without a pre-measurement mixing step.
- Statistical analysis, including two-tailed paired t-tests and multiple comparison corrections, was used to compare doubling times.
Main Results:
- Mixing did not significantly alter the biological interpretation of cell growth dynamics.
- Averaged across eight strains, doubling times between mixed and unmixed samples showed a difference of approximately 10%.
Conclusions:
- The mixing step prior to OD determination can be omitted in HTS workflows if a ~10% variability is acceptable.
- Simplifying HTS by removing the mixing step can enhance cost-effectiveness and process efficiency without compromising data accuracy.
