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Automated and online characterization of adherent cell culture growth in a microfabricated bioreactor.

Jaccard N, Macown RJ, Super A, Griffin LD, Veraitch FS, Szita N - J Lab Autom (2014)

Bottom Line: While suspension culture processes benefit from decades of development of instrumented bioreactors, adherent cultures are typically performed in static, noninstrumented flasks and well-plates.A machine learning-based algorithm enabled the specific detection of one cell type within a co-culture setting, such as human embryonic stem cells against the background of fibroblast cells.In addition, the algorithm did not confuse image artifacts resulting from microfabrication, such as scratches on surfaces, or dust particles, with cellular features.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemical Engineering, University College London, London, UK Centre for Mathematics and Physics in the Life Sciences and Experimental Biology, University College London, London, UK.

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Schematic of the key components required for automated culture and monitoring of adherent cells in a microfabricated bioreactor. (A) Flow is controlled either by modulating the head pressure in a bottle containing the culture media or via a syringe drive. Temperature control is achieved using an on-stage incubator that houses the microfabricated bioreactor as well as the fluidics. A motorized stage is used together with a piezo focus system for imaging. (B) Schematic of a typical monitoring loop for the system. Automation of the fluidics and the imaging system is achieved using a LabVIEW routine while automated image processing was done using MATLAB.
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fig1-2211068214529288: Schematic of the key components required for automated culture and monitoring of adherent cells in a microfabricated bioreactor. (A) Flow is controlled either by modulating the head pressure in a bottle containing the culture media or via a syringe drive. Temperature control is achieved using an on-stage incubator that houses the microfabricated bioreactor as well as the fluidics. A motorized stage is used together with a piezo focus system for imaging. (B) Schematic of a typical monitoring loop for the system. Automation of the fluidics and the imaging system is achieved using a LabVIEW routine while automated image processing was done using MATLAB.

Mentions: The use of a microfabricated device as both a culture and imaging chamber requires the following key components (Fig. 1A): flow control for culture medium exchange, environmental control to maintain optimal temperature for growth, and an imaging system for monitoring. The implementation of these three components depends on the monitoring strategy. Intermittent monitoring allows the transport of the microfabricated device from its controlled environment (e.g., incubator) to the microscope for image acquisition. In contrast, online monitoring is achieved by having the device permanently positioned on top of a microscope stage and thus requires a suitable setup to maintain optimal growth conditions. Online monitoring offers obvious advantages, such as high image sampling rates, flexible monitoring schedules, reduced contamination risks (no need to transfer to and from an incubator), and a higher degree of automation (and thus reduced user interaction). Its main limitation lies in the number of devices that can be monitored at once with one microscope. To demonstrate the applicability of our imaging and automation routines, we tested both monitoring concepts.


Automated and online characterization of adherent cell culture growth in a microfabricated bioreactor.

Jaccard N, Macown RJ, Super A, Griffin LD, Veraitch FS, Szita N - J Lab Autom (2014)

Schematic of the key components required for automated culture and monitoring of adherent cells in a microfabricated bioreactor. (A) Flow is controlled either by modulating the head pressure in a bottle containing the culture media or via a syringe drive. Temperature control is achieved using an on-stage incubator that houses the microfabricated bioreactor as well as the fluidics. A motorized stage is used together with a piezo focus system for imaging. (B) Schematic of a typical monitoring loop for the system. Automation of the fluidics and the imaging system is achieved using a LabVIEW routine while automated image processing was done using MATLAB.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2 - License 3
Show All Figures
getmorefigures.php?uid=PMC4230958&req=5

fig1-2211068214529288: Schematic of the key components required for automated culture and monitoring of adherent cells in a microfabricated bioreactor. (A) Flow is controlled either by modulating the head pressure in a bottle containing the culture media or via a syringe drive. Temperature control is achieved using an on-stage incubator that houses the microfabricated bioreactor as well as the fluidics. A motorized stage is used together with a piezo focus system for imaging. (B) Schematic of a typical monitoring loop for the system. Automation of the fluidics and the imaging system is achieved using a LabVIEW routine while automated image processing was done using MATLAB.
Mentions: The use of a microfabricated device as both a culture and imaging chamber requires the following key components (Fig. 1A): flow control for culture medium exchange, environmental control to maintain optimal temperature for growth, and an imaging system for monitoring. The implementation of these three components depends on the monitoring strategy. Intermittent monitoring allows the transport of the microfabricated device from its controlled environment (e.g., incubator) to the microscope for image acquisition. In contrast, online monitoring is achieved by having the device permanently positioned on top of a microscope stage and thus requires a suitable setup to maintain optimal growth conditions. Online monitoring offers obvious advantages, such as high image sampling rates, flexible monitoring schedules, reduced contamination risks (no need to transfer to and from an incubator), and a higher degree of automation (and thus reduced user interaction). Its main limitation lies in the number of devices that can be monitored at once with one microscope. To demonstrate the applicability of our imaging and automation routines, we tested both monitoring concepts.

Bottom Line: While suspension culture processes benefit from decades of development of instrumented bioreactors, adherent cultures are typically performed in static, noninstrumented flasks and well-plates.A machine learning-based algorithm enabled the specific detection of one cell type within a co-culture setting, such as human embryonic stem cells against the background of fibroblast cells.In addition, the algorithm did not confuse image artifacts resulting from microfabrication, such as scratches on surfaces, or dust particles, with cellular features.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemical Engineering, University College London, London, UK Centre for Mathematics and Physics in the Life Sciences and Experimental Biology, University College London, London, UK.

Show MeSH
Related in: MedlinePlus