Mixing is Dispensable for Optical Density-Based High-Throughput Growth Screening Assay in Fission Yeast

Lim, Kim Kiat; Chung, Jiunn Jye; Ma, Sha; Yen, Ching-Chiuan; Zhang, Louxin; Chen, Ee Sin

doi:10.3390/ijms27083410

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Mixing is Dispensable for Optical Density-Based High-Throughput Growth Screening Assay in Fission Yeast

by

Kim Kiat Lim

^1,*

,

Jiunn Jye Chung

²,

Sha Ma

²,

Ching-Chiuan Yen

²

,

Louxin Zhang

³ and

Ee Sin Chen

^1,4,5,6,*

¹

Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117596, Singapore

²

NUS Centre for Additive Manufacturing, National University of Singapore, Singapore 117575, Singapore

³

Department of Mathematics, Faculty of Science, National University of Singapore, Singapore 119076, Singapore

⁴

National University Health System (NUHS), Singapore 119228, Singapore

⁵

NUS Graduate School of Science & Engineering, National University of Singapore, Singapore 119077, Singapore

⁶

NUS Artificial Intelligence Institute, National University of Singapore, Singapore 119391, Singapore

^*

Authors to whom correspondence should be addressed.

Int. J. Mol. Sci. 2026, 27(8), 3410; https://doi.org/10.3390/ijms27083410

Submission received: 2 February 2026 / Revised: 6 April 2026 / Accepted: 7 April 2026 / Published: 10 April 2026

(This article belongs to the Special Issue Advances in Yeast Engineering and Stress Responses)

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Abstract

Optical density (OD)-based cell growth measurement is commonly used in high-throughput screening (HTS) during drug discovery or when deciphering the pharmaceutical mechanism of action. While resuspending the cells via a mixing step is often assumed to be necessary prior to OD measurement, its essentiality in HTS workflows has not been systematically verified. Here, through the measurement of the growth of several strains of the microbial yeast Schizosaccharomyces pombe cells, we compared the overall growth dynamics between samples that have been mixed and not mixed. Using statistical quantification by a two-tailed paired t-test followed by multiple comparison corrections, we concluded from the comparison of the doubling time of cells growing in the exponential phase that mixing did not significantly affect the biological interpretation compared to unmixed samples. Doubling time quantification between mixed and unmixed samples showed a difference of approximately 10% on average based on the assessment of the growth of eight strains. As such, if the experimental outcome can accommodate this level of variability, incorporating a mixing step before OD determination would not be necessary. These observations support the simplification of HTS processes, improving the cost efficacy and process efficiency of readouts, yet maintaining the accuracy of data acquisition.

Keywords: fission yeast; Schizosaccharomyces pombe; microtiter plate; growth rate; doubling time; exponential phase

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MDPI and ACS Style

Lim, K.K.; Chung, J.J.; Ma, S.; Yen, C.-C.; Zhang, L.; Chen, E.S. Mixing is Dispensable for Optical Density-Based High-Throughput Growth Screening Assay in Fission Yeast. Int. J. Mol. Sci. 2026, 27, 3410. https://doi.org/10.3390/ijms27083410

AMA Style

Lim KK, Chung JJ, Ma S, Yen C-C, Zhang L, Chen ES. Mixing is Dispensable for Optical Density-Based High-Throughput Growth Screening Assay in Fission Yeast. International Journal of Molecular Sciences. 2026; 27(8):3410. https://doi.org/10.3390/ijms27083410

Chicago/Turabian Style

Lim, Kim Kiat, Jiunn Jye Chung, Sha Ma, Ching-Chiuan Yen, Louxin Zhang, and Ee Sin Chen. 2026. "Mixing is Dispensable for Optical Density-Based High-Throughput Growth Screening Assay in Fission Yeast" International Journal of Molecular Sciences 27, no. 8: 3410. https://doi.org/10.3390/ijms27083410

APA Style

Lim, K. K., Chung, J. J., Ma, S., Yen, C.-C., Zhang, L., & Chen, E. S. (2026). Mixing is Dispensable for Optical Density-Based High-Throughput Growth Screening Assay in Fission Yeast. International Journal of Molecular Sciences, 27(8), 3410. https://doi.org/10.3390/ijms27083410

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Mixing is Dispensable for Optical Density-Based High-Throughput Growth Screening Assay in Fission Yeast

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