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Flow Cytometry

Flow Cytometry

The development of flow cytometry techniques began in 1934 with initial attempts by Andrew Moldavan, a bacteriologist who counted the cells in a flowing capillary system. Moldavan pumped cells through a capillary tube focused under a microscope for visualization. The invention of photometry allowed the measurement of differentially-stained cells, and Louis Kamentsky developed the first multiparameter flow cytometer in 1965 to identify and count the cancer cells in cervical tissue specimens.
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Assembly and Quantification of Co-Cultures Combining Heterotrophic Yeast with Phototrophic Sugar-Secreting Cyanobacteria

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Automated yeast cultivation control using a biosensor and flow cytometry.

Raquel Perruca Foncillas¹, Sara Magnusson¹, Basel Al-Rudainy²

¹Division of Applied Microbiology, Department of Chemistry, Lund University, SE-22100 Lund, Sweden.

Journal of Industrial Microbiology & Biotechnology

|October 18, 2024

Summary

This study presents an automated control system for microbial bioprocessing. It uses flow cytometry and biosensors to monitor yeast cells and maintain optimal conditions, even under stress from furfural.

Keywords:

automated real-time flow cytometry control strategy fermentation heterogeneity synthetic biology

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Area of Science:

Biotechnology and Bioprocessing
Microbial Physiology
Synthetic Biology

Background:

Effective microbial bioprocessing requires precise control over cellular performance and robustness.
Traditional monitoring methods often lack the sensitivity to detect subtle physiological changes.
Environmental stressors like furfural can negatively impact microbial cultivation efficiency.

Purpose of the Study:

To evaluate an automated fed-batch cultivation control system.
To utilize at-line flow cytometry with a redox biosensor for monitoring yeast cell physiology.
To assess the system's ability to maintain cellular fitness under furfural stress.

Main Methods:

Development of a fed-batch cultivation control system employing at-line flow cytometry.
Use of intact yeast cells with a fluorescent transcription factor-based redox biosensor (TRX2p-yEGFP).
Monitoring of the NADPH/NADP+ ratio in response to furfural exposure.

Main Results:

The control system successfully detected biosensor output and automatically adjusted furfural feed rate.
Physiological fitness of the yeast strain was maintained even at high furfural concentrations.
Single-cell measurements allowed for the monitoring of subpopulation dynamics, improving control precision.

Conclusions:

Automated control systems combining biosensors and flow cytometry offer robust microbial cultivation.
Leveraging intracellular properties as control inputs enhances system performance.
This approach holds significant potential for optimizing industrial bioprocessing applications.