Summary of Contents for SBI CGQ
- Page 1 CGQ user guide A handbook for cell density monitoring in shake flask and bioreactor applications Revision 2...
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Page 3: Table Of Contents
Table of contents Introduction and general considerations ........5 The CGQ system – an overview .............. 7 Absolute maximum ratings ..............8 Recommended operating conditions ............. 9 Warnings ....................11 Declarations and certificates ..............13 CE conformity ..................13 WEEE conformity ................14 FCC compliance ................. - Page 4 Optical cell density measurements ............34 Challenges in shaken and stirred vessels ........... 37 Correlation between OD and the CGQ signal ........39 Parameters that influence the CGQ measurements ......41 Contact ..................48 CGQ experiment checklist ..............49...
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Page 5: Introduction And General Considerations
To ensure that this user guide provides all information you need during your work with the CGQ, we at SBI are reliant on your feedback. Do not hesitate to contact us to share your ideas regarding errors, missing information, or incomprehensibilities so that we can improve this document and keep it up to date with your requirements. - Page 6 CGQ user guide revisions: Revision 0 02.09.2022 Initial document release Revision 1 19.04.2023 Updated illustration of BioR LEDs Revision 2 21.11.2023 Updated SBI North America address...
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Page 7: The Cgq System - An Overview
The CGQ system – an overview The CGQ is an analytical laboratory device that is primarily intended to be used for noninvasive cell density monitoring in real-time, especially in shake flasks (CGQ) and bioreactor (CGQ BioR) applications. However, even completely different applications requiring the real-time analysis of particle concentrations can be addressed by the CGQ system. -
Page 8: Absolute Maximum Ratings
While it is not guaranteed to work at 75°C, the CGQ will do that in most expectable cases. Operating the CGQ above 50°C requires special darkening covers,... -
Page 9: Recommended Operating Conditions
Recommended operating conditions 10 – 50 °C Temperature (Ensure to let the CGQ Sensors adjust to the operating temperature for 30 min.) 0 – 80% (non-condensing) Humidity (relative) Shake flask filling volume 10 – 15% optimal range 5 – 25% good range 2 –... - Page 10 Constant ambient light can be compensated efficiently by the CGQ. Strong changes of the ambient light intensity (e.g. from absolute darkness at night to bright daylight at a sunny mounting position) may be visible as step-like artifacts in the measurement data.
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Page 11: Warnings
CGQ and any other electric device around. Do not look into the beam of any of the CGQ Sensor LEDs! Their emitted light is of high intensity and might damage your eye or retina. Wear protective eye wear. Avoid direct eye and skin exposure to the CGQ Sensor LEDs! Always keep a safety distance of >1 m to any active CGQ Sensor LED. - Page 12 Do not use inorganic or organic acids and bases, organic solvents or detergents to clean the CGQ! Some solvents or detergents might be allowed for cleaning, but you should only use those being mentioned in the user guide. Any kind of opening, manipulating or copying CGQ devices as...
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Page 13: Declarations And Certificates
The aquila biolabs GmbH, Arnold-Sommerfeld-Ring 2, 52499 Baesweiler, Germany, herewith declares under its sole responsibility that all devices and equipment being part of the CGQ system and being manufactured by the aquila biolabs GmbH are in conformity with the Council Directives as described in EN... -
Page 14: Weee Conformity
The aquila biolabs GmbH, Arnold-Sommerfeld-Ring 2, 52499 Baesweiler, Germany, herewith declares compliance of all electronic components of the CGQ system with the Council Directive 2012/19/EU. Electronic components may contain various hazardous substances that could possibly exhibit negative impacts on your health and the environment. In order... -
Page 15: Fcc Compliance
FCC compliance This is a Class A product. This device complies with Part 15 of the FCC Rules. Operation is subject to the following two conditions: (1) this device may not cause harmful interference, and (2) this device must accept any interference received, including interference that may cause undesired operation. -
Page 16: Installation
Installation The CGQ system can be used for online biomass monitoring in shake flasks or bioreactor vessels. The main difference is the sensor type: CGQ Sensor for shake flasks, CGQ BioR for bioreactors. The following section explains the components and installation process for shake flasks. Refer to page 27 for bioreactors. -
Page 17: Installation On A Shaking Tray With Screw Threads
CGQ sensor at its side. The side-measurement is required due to the thick flask bottom. The CGQ Hub fits on any kind of shaking tray with screw threads or sticky pad. The following sections guide you through the installation of all system components step-by-step. - Page 18 Hub, just move it over the tray until at least two screwing threads are visible through the elongated CGQ Hub bottom plate holes. Then use appropriate screws that fit your tray’s screw threads to fix the CGQ Hub on the tray. Avoid the usage of countersunk head screws, as they might destroy the CGQ Hub bottom plate when being screwed too tightly.
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Page 19: Installation On A Shaking Tray With Sticky Mat
Figure 4, with the ports of the Hub facing towards the inside of the shaker. Push down the CGQ Hub to make sure that it is in good contact with the sticky mat. At higher shaking frequencies (> 200 rpm), it is recommended to request an alternative bottom plate from SBI, which allows to mount the CGQ Hub on its backside. - Page 20 CGQ Hub and the Sticky Mat/Clamp Adapters by pulling slowly, then wait until the component detaches from the sticky mat. Do not use liquids for the removal of the CGQ Hub from a sticky mat shaker. The liquid might damage the CGQ Hub electronics...
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Page 21: Mounting A Cgq Sensor
Mounting a CGQ Sensor The CGQ Sensors fit into Infors/Lauber spring clamps for 100 ml flasks. For larger flasks, use adapters as shown in Figure 5. Figure 5: Assembly of CGQ Sensor with Sensor Adapter for 250 ml flask. Always mount the CGQ Sensor according to the following instructions. - Page 22 Figure 8: Mounting the CGQ in a Sensor Adapter – Step 1. Tighten the handle on the adapter to secure the CGQ Sensor in place.
- Page 23 Make sure that the CGQ Sensor cable is not pinched between CGQ Sensor and spring clamp. Otherwise, the device might be damaged during shaking.
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Page 24: Mounting The Shake Flask
Insert the flask into the spring clamp as usual. Turn the shake flask around its vertical axis until neither the white labeling area nor potentially present baffles are located above the sensor array. Figure 11: Mounting the shake flask on top of the CGQ Sensor. -
Page 25: Cable Management Within The Shaker
Always mount the CGQ Hub in the shaker, on the tray on which the connected sensors are installed. The CGQ Hub connectors have to face the inner side of the shaker. - Page 26 Figure 12. Use the cable clamp array from your CGQ package as shown in Figure 13 and resize it according to your requirements. It can either be screwed or glued to position.
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Page 27: Installing The Bior On A Bioreactor
Figure 15: The BioR Sensor with elastic foam and Velcro belt. The CGQ BioR is equipped with a thick layer of elastic foam and with a mounting belt. Both together allow you to install the CGQ BioR at almost any bioreactor in your lab. - Page 28 Figure 16: Exemplary setups of BioR Sensors mounted on bioreactors, mounting the belt around the complete bioreactor setup, around the vessel, around the vessel and rods and around two rods. Do not mount the BioR Sensor on top of marks or labels on the vessel, as this may negatively influence the data quality.
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Page 29: Connection To Dots Software
BioR Sensor cables. Connect the BioR Sensor to any port of the CGQ Hub. Connect the power cable and the USB cable on top of the CGQ Hub. Take care of proper cable routing. Connection to DOTS Software The latest firmware should be installed on CGQ Sensors and BioR Sensors. - Page 30 LED should start glowing and the CGQ should enumerate as COM device on your computer. Try to find a good way to guide the cables out of the shaker. The goal must be to reduce the cables’ shearing forces during shaking as much as possible, thus ensuring an optimal contact between cable plug and Hub socket.
- Page 31 3. Mount the shake flask or inoculate your bioreactor Mount your inoculated shake flask on top of the sensor plate into the spring clamp (pp. 24). If you are working with a CGQ BioR, inoculate and prepare your bioreactor.
- Page 32 Wrong positioning might cause measurement artifacts (pp. 41). Do not mount the CGQ BioR on top of marks or labels on the vessel, as this may negatively influence the data quality. 4. Start your cultivation All hardware related works have been done right now, so you can close and start your shaker.
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Page 33: Cleaning And Disinfection
PC, to prevent damages to the electronics, to connected devices and to your health. Ensure that all CGQ devices and cables are completely dry after disinfection, before you reconnect them to each other and to the power supply and USB. -
Page 34: Cgq's Basic Measurement Principles
CGQ’s basic measurement principles Optical cell density measurements The CGQ’s technique for noninvasive cell density monitoring in shake flasks is based on the principle of light scattering. Basically, each measurement consists of a sequence of three major steps. (1) Light is irradiated by a LED into the fermentation broth through the transparent vessel wall. - Page 35 Please also refer to the recommended operating conditions (pp. 9) While the standard CGQ Sensor peak wavelength is 521 nm, the BioR Sensors contain an additional infrared LED (940 nm). The probability for infrared photons...
- Page 36 Position 1: 521 nm Deep Blue True Yellow Amber Red Hyper IR I IR II Blue Green Figure 18: Standard LED colors at the CGQ and BioR measurement positions and LED colors available for wavelength customization of CGQ and BioR Sensors.
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Page 37: Challenges In Shaken And Stirred Vessels
Using an array of LEDs and photodiodes, each CGQ Sensor can adaptively measure the backscattering intensity at different radial positions, thus accounting for a broad variety of liquid distributions in shake flask fermentations. - Page 38 The CGQ does not treat the moving-liquid-induced backscattering signal fluctuations as noise, but as a source of valuable information. For determining one cell density value, the CGQ Sensor collects about one million of single backscattering intensities per second; thereby creating a series of data points that combines information regarding cell density and liquid distribution on a microseconds-resolution.
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Page 39: Correlation Between Od And The Cgq Signal
Lambert-Beer’s law. As described above, the CGQ measures backscattered light intensities. While scattering signals can be used to measure particle concentrations over several orders of magnitude, it must be noticed that there is no physical law to describe... - Page 40 The DOTS Software implements methods to correlate offline cell density data such as OD or Cell Dry Weight with the CGQ’s backscattered light signal. Additional information is provided in the DOTS Software User Guide.
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Page 41: Parameters That Influence The Cgq Measurements
However, this sensitivity means that also changes in the optical environment will be detected by the CGQ. With the ambient light compensating CGQ and BioR Sensors it is possible to monitor growth without the use of covers. - Page 42 100% cumulative distribution -12% deviation from average Figure 21: Exemplary CGQ signal differences of a set of 24 shake flasks (aluminum cap, 250 ml), measured at OD = 4.2, 250 rpm, 25mm shaking diameter, 25 ml filling volume. As long as the flasks’ positions or orientations are not changed during...
- Page 43 In some cases, it may be favorable to preselect a set of highly similar flask, based on measuring their scattering signal on the same CGQ Sensor, at the same cell density and at equal shaking conditions. Shake flasks and bioreactors are not a blank cuvette! While cuvettes typically exhibit blank surfaces, shake flasks and bioreactors are usually covered by volume markings, engravings and labeling areas.
- Page 44 Generally, this is not problematic for the cell density measurement; however different materials might have different effects on the CGQ signal. As long as the material is at least partially transparent for visible light, the CGQ measurement principle will work, but care must be taken about some limitations.
- Page 45 CGQ signal (refer to pp. 37). The CGQ’s data analysis algorithms are made and continuously improved (via firmware updates) to eliminate as many of these signal-influencing fluidic parameters as possible.
- Page 46 Even slight changes in the clamp deformation can introduce considerable differences in the relative positioning of shake flask and CGQ Sensor, thus influencing the effective scattering liquid volume in the sensor’s field of view.
- Page 47 CGQ’s scattering signal and may therefore negatively influence the flask-to-flask reproducibility and comparability of similar CGQ measurements. In order to avoid this effect, it is strongly recommended to check the symmetry of all clamps each time before mounting a shake flask for CGQ measurements.
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Page 48: Contact
Newsletter https://www.scientificbio.com/subscription-center subsciption Information on new Software versions, Device firmware updates, new products Email insights@scientificbio.com Headquarters SBI North America SBI Europe Represented by: aquila biolabs GmbH 201 Penn Center Blvd Arnold-Sommerfeld-Ring 2 Pittsburgh, PA 15235 52499 Baesweiler, Germany +1 413-230-8463... -
Page 49: Cgq Experiment Checklist
CGQ experiment checklist Is the CGQ Hub connected to the power supply (green LED glowing)? Is the CGQ Hub connected to PC and DOTS Software via USB (CGQ Hub visible and appears as connected in Device list)? Are the CGQ USB and power cables routed with sufficient play within...
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