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Foam Formation in Shake Flasks and Its Consequences.

David Vonester1, Kyra Hoffmann1, Thomas Palmen1

  • 1AVT - Biochemical Engineering, RWTH Aachen University Aachen Germany.

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|October 24, 2025
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Summary
This summary is machine-generated.

Foam formation in shake flasks, especially non-baffled ones, can unexpectedly increase oxygen transfer capacity (OTRmax) and reduce experimental reproducibility. This study investigates the causes and impacts of foaming in shake flask cultivations.

Keywords:
foam formationmaximum oxygen transfer capacityonline viscosity measurementout‐of‐phase phenomenonshake flask

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Area of Science:

  • Biotechnology and bioprocess engineering
  • Microbial cultivation and fermentation technology
  • Process analytical technology

Background:

  • Foam formation is a known issue in stirred tank fermentations but often overlooked in shake flask cultivations.
  • Non-baffled shake flasks are generally assumed to prevent foaming, but this is not always the case under specific conditions.
  • Understanding foam dynamics in shake flasks is crucial for accurate experimental interpretation and process optimization.

Purpose of the Study:

  • To investigate the phenomena of foam formation in shake flasks, particularly non-baffled systems.
  • To determine the impact of foaming on maximum oxygen transfer capacity (OTRmax).
  • To assess the effect of foaming on the reproducibility of shake flask experiments.

Main Methods:

  • Experimental investigation of foam formation in shake flasks under various process conditions.
  • Measurement of oxygen transfer capacity (OTRmax) in the presence and absence of foam.
  • Analysis of factors inducing foaming, including vessel internals and cultivation phenomena.

Main Results:

  • Foaming in shake flasks can elevate the maximum oxygen transfer capacity (OTRmax) by up to threefold.
  • This increase in OTRmax significantly alters process conditions, negatively impacting experimental reproducibility.
  • Foaming can be induced by vessel components like baffles or sensor spots and is linked to the out-of-phase phenomenon, especially at elevated viscosities.

Conclusions:

  • Foam formation in shake flasks is a significant factor that alters process conditions and must be considered.
  • The observed increase in OTRmax due to foaming can lead to misleading results and reduced reproducibility.
  • Strategies to ensure in-phase cultivation and unimpeded liquid flow are recommended to mitigate foaming in shake flasks.