Make Accurate, Real-Time Decisions with Flow Cytometry

Flow cytometry has traditionally been limited to high-end research labs due to its cost and complexity. BactoBox makes flow cytometry accessible for production environments, offering a novel in-process control method to reduce process variability, optimize key decision points, and improve yields with rapid, precise bacterial enumeration.

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Why Flow Cytometry Can Improve Your Production

Traditional microbial monitoring methods, such as plating and optical density (OD) measurements, introduce delays and uncertainties that can compromise production efficiency. Flow cytometry provides a faster, more precise way to monitor bacterial concentrations in real-time, enabling better decision-making at critical process points.

By integrating BactoBox and flow cytometry in your production, you can:

Track growth dynamics with precision – Detect even small fluctuations in growth rates and perform corrective actions.
Maximize process yield – Harvest at the optimal timepoint to ensure peak productivity.
Ensure batch-to-batch consistency – Standardize inoculum concentrations for reproducible outcomes.
Detect early signs of cell death – Identify changes in bacterial viability before they impact production.

These capabilities allow for a more efficient, higher-yield production process with improved control over microbial dynamics. In our experience, OD600 readings are often skewed or delayed, surprising many users who switch to flow cytometry for real-time insights.

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A Source of Variability: How Inoculum Size Affects Growth Translation

Accurately controlling bacterial inoculum size is critical for maintaining consistency in fermentation processes. Small variations in initial cell concentration can significantly impact growth rates, time-to-target concentration, and overall process efficiency. With real-time bacterial enumeration, manufacturers can eliminate a major source of variability by ensuring precise inoculum control at every stage of the seed train.

The graph above illustrates how inoculum size influences the growth of E. coli. Cultures were inoculated with 1×10³, 1×10⁵, and 1×10⁷ cells/mL, with inoculum sizes precisely adjusted using BactoBox. All cultures originated from the same starter culture. As expected, the time required to reach 1×10⁹ cells/mL varied significantly based on the initial inoculum concentration, ranging from 3.8 hours to 9.0 hours.

Without precise enumeration, inoculation steps in a seed train can introduce significant variability. Many manufacturers rely on OD600 measurements to estimate inoculum concentration, but OD600 is an indirect proxy that is influenced by cell morphology, aggregation, and media composition. The uncertainty of OD600-based measurements compounds over multiple inoculation steps, leading to unpredictable process outcomes.

Ensure Consistency with BactoBox

With BactoBox, you can directly measure bacterial concentration in minutes, ensuring that each inoculation step starts with the correct number of cells. By replacing OD600 estimates with direct cell counts, you can:

Minimize batch-to-batch variation
Reduce process variability and time-to-target concentration
Improve consistency across fermentation runs

Optimize your microbial production with real-time bacterial enumeration. Contact us to learn how BactoBox can improve your process efficiency.

Using BactoBox for Industrial Microbial Production - FAQ

Find answers to the most frequently asked questions when using BactoBox in a industrial microbial production setting.

How easy is it to use BactoBox?

BactoBox is designed for ease of use, enabling rapid bacterial enumeration with minimal training. The measurement process consists of three simple steps: homogenization, dilution, and measurement. Our experience is that we can train anyone, and for proficient users, sample preparation and measurement can be completed in under 5 minutes.

Can BactoBox perform online or at-line sampling?

No, BactoBox does not support online or at-line sampling. It operates using a grab sample approach, making it ideal for evaluating key decision points throughout the fermentation process. This allows users to monitor bacterial concentrations at critical stages without the complexity of continuous monitoring systems.

Can I use BactoBox in media that contain non-celluar particles?

Yes, but a one-time gating customization is required to ensure non-cellular particles do not affect bacterial counts. This customization is specific to your bacterial genera and allows accurate enumeration in complex media.

Can BactoBox count both live and dead cells?

With customization, BactoBox can detect the onset of cell death. It shows a trend correlation to plate counting, but results will not match one-to-one since CFUs cannot detect viable-but-non-culturable cells, and some dead cells may still have an intact membrane. BactoBox is a useful tool for monitoring when cell death begins in your process.