Applied Microbiology and Biotechnology

, Volume 102, Issue 10, pp 4279–4289 | Cite as

Practices of shake-flask culture and advances in monitoring CO2 and O2

  • Masato Takahashi
  • Hideki Aoyagi


About 85 years have passed since the shaking culture was devised. Since then, various monitoring devices have been developed to measure culture parameters. O2 consumed and CO2 produced by the respiration of cells in shaking cultures are of paramount importance due to their presence in both the culture broth and headspace of shake flask. Monitoring in situ conditions during shake-flask culture is useful for analysing the behaviour of O2 and CO2, which interact according to Henry’s law, and is more convenient than conventional sampling that requires interruption of shaking. In situ monitoring devices for shake-flask cultures are classified as direct or the recently developed bypass type. It is important to understand the characteristics of each type along with their unintended effect on shake-flask cultures, in order to improve the existing devices and culture conditions. Technical developments in the bypass monitoring devices are strongly desired in the future. It is also necessary to understand the mechanism underlying conventional shake-flask culture. The existing shaking culture methodology can be expanded into next-generation shake-flask cultures constituting a novel culture environment through a judicious selection of monitoring devices depending on the intended purpose of shake-flask culture. Construction and sharing the databases compatible with the various types of the monitoring devices and measurement instruments adapted for shaking culture can provide a valuable resource for broadening the application of cells with shake-flask culture.


Bioprocess monitoring Carbon dioxide Monitoring device Oxygen Shake-flask culture 



The manuscript has been edited by native English-speaking professional editors from Editage, a division of Cactus Communications.

Authors’ information

Masato Takahashi (postdoctoral researcher; research fellow of the Japan Society for the Promotion of Science). Faculty of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Ibaraki 305-8572, Japan

Hideki Aoyagi (professor and principal investigator). Division of Life Sciences and Bioengineering, Graduate School of Life and Environmental Sciences and Faculty of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Ibaraki 305-8572, Japan

Funding information

This work was supported by a grant-in-aid for Japan Society for the Promotion of Science (JSPS) fellows (grant number 16J00800). This work was a funded by a JSPS KAKENHI (grant number 15H04569), JSPS KAKENHI Challenging Research [Exploratory] (grant number 17K19218), and the Sumitomo Electric Industries Group Corporate Social Responsibility Foundation.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.


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Authors and Affiliations

  1. 1.Faculty of Life and Environmental SciencesUniversity of TsukubaTsukubaJapan

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