Pre-sterilized 48 well microplates or 32 well (+16 reservoir wells) microfluidic plates are available in classic round well or flower-shaped geometry which provides a baffle-like effect for improved oxygen transfer and mixing. Plates with microfluidic technology have a bottom containing microchannels that connect two reservoir wells of a column with each of the four cultivation wells underneath. Every cultivation well of a column can receive up to two actuators from their corresponding reservoir wells and pressure-controlled microvalves enable individual control of these actuators for each cultivation well at nanoliter scale. This allows the pH value to be controlled for single wells by various acids and alkalis, as well as the continuous addition of nutrient solutions, such as glucose or glycerol.
Gassing modules facilitate mixing of gases to set oxygen or carbon dioxide levels. Setpoints of gas concentrations can be changed over the cultivation, yet only one gassing module can be used during a cultivation experiment. The gas mixture is optionally channeled through a water bottle for humidification in order to reduce evaporation of the cultures. Gas is sparged through gassing lids covering the microplate allowing continuous gas exchange and low gas consumption. The gassing lid seals the microplate air-tightly from the environment, which makes it possible to also run anaerobic cultivations even though the BioLector XT instrument itself is positioned in usual atmosphere. Beware that microfluidic technology requires the gassing lid to be added right before starting the experiment and Ginkgo Bioworks does not have the anaerobic environment required to facilitate very strict anaerobes but less strict anaerobic cultures have successfully been cultivated already.
The optical measurements performed by the instrument are realized by using a wide range of filter modules for the detection of scattered and fluorescent light. Up to six different filter modules can be used during an experiment to measure in sequence. Optodes embedded in the well bottom allow for detection of pH value and DO concentration in the liquid phase.
General ranges:
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Volume
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800 - 2400 µL (depending on plate and agitation)
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Temperature
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10 – 50 °C (minimum temperature 8 °C below ambient temperature)
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Gas flow
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5-50 mL/min (variable flow rate is programmable up front)
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Agitation
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100 – 1500 rpm (depending on plate and used volumes; 3 mm diameter)
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pH
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pH 4.2 – 7.5 (depending on plate optode)
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DO
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0 – 100% oxygen saturation (100% corresponding to the DO level reached while gassing with 100% O2 without O2 consumption)
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Scattered Light Measurement
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Resolution > 50 NTU, at densities higher than 500 NTU: 10 % of measured value.
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Wavelengths
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365 nm–800 nm (filter modules with specific wavelengths)
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kLa values
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25 – 600 1/h
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Present modules:
o Biomass (scattered light)
o pH (HP8 optode, range 4.4 - 7.8, accurate between 5.1 - 7.1)
o pH (LG1 optode, range 3.7 - 8.0, accurate between 4.7 - 7.0)
o pH low (pH51 optode, range 3.6 - 6.5, accurate between 4.2 - 6.0)
o DO (Pst3 optode)
o DO (RedFlash optode)
o Riboflavin
o O2 Up-Regulation Module, 21-100%
o O2 Down-Regulation Module, 1-21%
o CO2 Up-Regulation Module, 0-12%
o Anaerobic Module (N2 or pre-mixed gas)
- Microfluidic Module (two-sided pH control, one-sided pH control plus feeding or two feeds).