MitoPedia: O2k hardware

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MitoPedia: O2k hardware

Template NextGen-O2k.jpg
O2k-Open Support


MitoPedia O2k and high-resolution respirometry: O2k-Open Support 

MitoPedia: O2k hardware is a component of O2k-Open Support.
It lists hardware-related topics and the O2k-Modules and main components of the O2k system.


See also
» Complete list of O2k components O2k-Products


TermAbbreviationDescription
Barometric pressurepb [Pa]Barometric pressure, pb, is an important variable measured for calibration of oxygen sensors in solutions equilibrated with air. The atm-standard pressure (1 atm = 760 mmHg = 101.325 kPa) has been replaced by the SI standard pressure of 100 kPa. The partial pressure of oxygen, pO2, in air is a function of barometric pressure, which changes with altitude and locally with weather conditions. The partial oxygen pressure declines by 12% to 14% per 1,000 m up to 6,000 m altitude, and by 15% to 17% per 1,000 m between 6,000 and 9,000 m altitude. The O2k-Barometric Pressure Transducer is built into the Oroboros O2k as a basis for accurate air calibrations in high-resolution respirometry. For highest-level accuracy of calculation of oxygen pressure, it is recommended to compare at regular intervals the barometric pressure recording provided by the O2k with a calibrated barometric pressure recording at an identical time point and identical altitude. The concept of gas pressure or barometric pressure can be related to the generalized concept of isomorphic pressure.
Block temperatureThe block temperature of the Oroboros O2k is the continuously measured temperature of the copper block, housing the two glass chambers of the O2k. The block temperature is recorded by DatLab as one of the O2k system channels.
ChannelF7» See O2k signals and output
FluorometryFluorometry (or fluorimetry) is the general term given to the method of measuring the fluorescent emission of a substance following excitation by light at a shorter wavelength.
GainThe gain is an amplification factor applied to an input signal to increase the output signal.
High-resolution respirometryHRR
O2k-FluoRespirometer

High-resolution respirometry, HRR, is the state-of-the-art approach in mitochondria and cell research to measure respiration in various types of mitochondrial preparations and living cells combined with MultiSensor modules. Mitochondrial function and dysfunction have gained an increasing interest over the past years, reflecting growing awareness of the fact that mitochondria play a pivotal role in human health and disease. Combining instrumental accuracy and reliability with versatility of applicable protocols - allowing practically unlimited addition and combination of substrates, inhibitors and uncouplers - mitochondrial respiratory pathways may be analyzed in detail to evaluate even minor alterations in respiratory pathway control and/or capacity. The most advanced way to analyze mitochondrial function is by means of high-resolution respirometry with the Oroboros O2k. The O2k is a sole source apparatus, with no other available instrument meeting its specifications for high-resolution respirometry. Substrate-uncoupler-inhibitor titration (SUIT) protocols allow the diagnosis of numerous mitochondrial pathway and coupling defects in a single respirometric assay. Technologically, HRR is based on the Oroboros O2k, combining optimized chamber design, application of oxygen-tight materials, electrochemical sensors, Peltier-temperature control, and specially developed software features (DatLab) to obtain the unique sensitive and quantitative resolution of oxygen concentration and oxygen flux, with both, a closed-chamber or open-chamber mode of operation (TIP2k). Standardized calibration of the polarographic oxygen sensor (static sensor calibration), calibration of the sensor response time (dynamic sensor calibration), and evaluation of instrumental background oxygen flux (systemic flux compensation) provide the experimental basis for high accuracy of quantitative results and quality control in HRR.

HRR can be extended for MultiSensor analysis by using the O2k-FluoRespirometer. Smart Fluo-Sensors are integrated into the O2k to measure simultaneously fluorometric signals using specific fluorophores. Potentiometric modules are available with ion selective electrodes (pH, TPP+). The NextGen-O2k is the all-in-one device including the Q-redox sensor and a PhotoBiology (PB) module.
Inside the O2kA glance inside the Oroboros O2k
Light-emitting diodeLEDA light-emitting diode (LED) is a light source (semiconductor), used in many every-day applications and specifically in fluorometry. LEDs are available for specific spectral ranges across wavelengths in the visible, ultraviolet, and infrared range.
O2kO2kO2k - Oroboros O2k: the modular system for high-resolution respirometry.
O2k repairO2k repair of defective hardware may require replacement of spare parts. Some electronic or mechanical defects may be solved only by repair of the O2k in the electronics workshop of Oroboros Instruments, e.g., a defective Peltier unit (temperature control).
O2k series
The serial number of each O2k is shown on a sticker at the rear of the O2k.
The O2k series is specified as the capital letter in the O2k serial number of the Oroboros O2k. A serial number G-#### or H-#### denotes an Oxygraph from the G or H series, while A-#### denotes an O2k from the A series. With DatLab running real-time connected to the O2k, the serial number of the currently connected O2k is displayed: (1) in the right corner of the status line, besides the DatLab version number (bottom), and (2) in windows O2k control [F7] and O2k configuration.
O2k signals and outputThree electronic channel types are available in the O2k-MultiSensor system. All channels are available twofold (dual-data), for O2k-Chambers A (left) and B (right), based on numerical signals sent at a fixed data sampling time interval (default: 2 s; range 0.2 s to >10 s).
O2k-Chamber MitoPediaO2k-Chamber: 16 mm inner diameter, Duran glass polished, with standard operation volume (V) of 2 cm3 (2 mL). Two units of this item are standard components mounted in the main unit of the O2k-FluoRespirometer.
O2k-Fluo LED2-ModuleThe O2k-Fluo LED2-Module is a component of the O2k-Fluorometer (O2k-Series D to G). It is an amperometric add-on module to the O2k-Core (O2k-Series D to G), adding a new dimension to high-resolution respirometry. Optical sensors are inserted through the front window of the O2k-glass chambers, for measurement of hydrogen peroxide production (Amplex UltraRed), ATP production (Magnesium green), mt-membrane potential (Safranin, TMRM), Ca2+ (Calcium green), and numerous other applications open for O2k-user innovation. The O2k-Fluo LED2-Module consists of optical sensors for both O2k-Chambers (LEDs for green and blue excitation), optical filters and the Fluorescence-Control Unit for regulation of light intensity and data input into the O2k-Main Unit (O2k-Series D to G). It includes the updated DatLab software.
O2k-Fluo Smart-ModuleThe O2k-Fluo Smart-Module is a component of the O2k-FluoRespirometer(O2k-Series H). It is an amperometric add-on module to the O2k-Respirometer, adding a new dimension to high-resolution respirometry. Optical sensors are inserted through the front window of the O2k-glass chambers, for measurement of hydrogen peroxide production (Amplex UltraRed), ATP production (Magnesium green), mt-membrane potential (Safranin, TMRM), Ca2+ (Calcium green), and numerous other applications open for O2k-user innovation. Current DatLab version is included in the delivery of this module.
O2k-FluoRespirometerThe Oroboros O2k-FluoRespirometer (O2k-Series H) - the experimental system complete for high-resolution respirometry (HRR), including fluorometry, the TIP2k and the O2k-sV-Module allowing simultaneous monitoring of oxygen consumption together with either ROS production (AmR), mt-membrane potential (TMRM, Safranin and Rhodamine 123), Ca2+ (CaG) or ATP production (MgG).

The O2k-FluoRespirometer supports all add-on O2k-Modules: O2k-TPP+ ISE-Module, O2k-pH ISE-Module, O2k-NO Amp-Module, enabling measurement of mt-membrane potential with ion sensitive electrodes (ISE for TPP+ or TPMP+ ) or pH.


Parameters:

  • oxygen consumption
  • ROS production
  • ATP production
  • Ca2+
  • mitochondrial membrane potential


Features of O2k-Series H:

  • Fluo-Control Unit integrated into O2k-Main Unit.
  • Smart Fluo-Sensors with pre-calibrated light intensities for direct input into DatLab 7
  • Added: spare OroboPOS
  • DL-Protocols as real-time guides for titrations in DatLab 7, context-sensitive help. Further details » MitoPedia: DatLab.
  • MiR05-Kit
  • MitoKit-CII


The O2k is a sole source apparatus with no other instruments meeting its specifications.
O2k-Fluorometer Series GO2k-Fluorometer Series G (up to 2017-July) - the experimental system complete for high-resolution respirometry (HRR) combined with fluorometry. The O2k-Fluorometer includes the O2k-Core, O2k-Fluo LED2-Module and TIP2k, and supports all other add-on O2k-Modules of the Oroboros O2k. The O2k is a sole source apparatus with no other instruments meeting its test experiments on O2k-Specifications.
O2k-Innovation descriptionO2kThe NextGen-O2k (NextGeneration O2k by Oroboros Instruments) is in development, building upon and extending the concept of the O2k-Fluorometer elaborated within the K-Regio Project MitoCom O2k-Fluorometer (2011-2014). To broaden the specifications of the O2k-FluoRespirometer, the NextGen-O2k will combine high-resolution respirometry with a Q-Module for measurements of Q-redox state and a PB-module to study PhotoBiology (including photosynthesis). It is our ultimate goal to further extend the NextGen-O2k with O2k-Spectrofluorometry (including ratiometric measurements), NADH autofluorescence, and O2k-Spectrophotometry (allowing for simultaneous measurement of redox states of the cytochromes aa3, b, and c). The Oroboros NextGen-O2k will have integrated optical elements beyond the modular options of the presently available Oroboros O2k.
O2k-NO Amp-ModuleO2k-NO Amp-Module: 2 black PEEK stoppers for amperometric O2k-MultiSensor application (NO, H2S, H2O2); 1 additional hole (2.3 mm); central titration port; conical bottom; with Volume-Calibration Rings (A and B); 2 O-rings mounted on each stopper, with 16 spare O-rings (O-ring\Viton\12.5x1 mm; two boxes). The NO sensor is not included.
O2k-Peltier Temperature ControlO2k-Peltier Temperature Control: Built-in electronic thermostat controlling temperature for two O2k-Chambers in the range of 4 to 47 °C; ±0.002 °C (at room temperature). Continuous recording of the O2k-Copper Block temperature with DatLab. Temperature change from 20 to 30 °C within 15 min; cooling from 30 to 20 °C within 20 min. Integral component of the O2k-Main Unit. The electronic temperature control of the O2k replaced the conventional water jacket.
O2k-SpecificationsO2k versus multiwell respirometer: O2k stands for Oroboros O2k and high-resolution respirometry, meeting powerful quality criteria securing high output and pioneering state-of-the-art comprehensive OXPHOS analysis of substrate control and coupling control of mitochondrial function. 'High throughput' stands for disposable multiwell systems - expensive, with limited scope and extremely high running costs. In respirometry, high throughput is not equivalent to high output. If you’re using a biased instrument, it doesn’t matter how many measurements you take – you’re aiming at the wrong target (Silver 2012 Penguin Press).
O2k-TPP+ ISE-ModuleO2k-TPP+ ISE-Module: 2 potentiometric ion-selective electrodes (ISE), ISE-Service Kit and 2 PEEK-Stoppers, supported by the Oroboros O2k (O2k-Series D and higher). It also includes the newest DatLab Version. The ISE module is designed with replaceable membranes making it possible to measure different ions such as TPP+ or Ca2+ with the same electrode housing1. The main application of the O2k-TPP+ ISE-Module is measurement of mitochondrial membrane potential with tetraphenylphosphonium (TPP+) or TPMP+. 1 There are other ion selective membranes available (e.g., from Sigma Aldrich), for a wide variety of ionic species. If cut to a circle with a diameter of 4 mm these membranes can be used in the ISE system. Many ions may also be detected by fluorimetric based methods.
O2k-pH ISE-ModuleO2k-pH ISE-Module: two pH electrodes and reference electrodes and accessories, two black PEEK stoppers, supported by the Oroboros O2k. It also includes the newest DatLab version. Quality control: Stability of Oroboros pH electrodes is tested for drift <15 µpH·s-1·mL-1.
O2k-sV-ModuleO2k-sV-Module: Now included in all O2k-Packages, comprised of Duran glass chambers of 12 mm inner diameter specifically developed to perform high-resolution respirometry with reduced amounts of biological sample, and all components necessary for smaller operation volume (V) of 0.5 cm3 (0.5 mL). Current DatLab version is included in the delivery of this revolutionary module.

The O2k-sV-Module is compatible with all O2k-Series and it is designed specifically for respirometry using small amounts of sample.

The O2k-sV-Module cannot be used for simultaneous measurement of multiple parameters in the O2k-FluoRespirometer or in combination with the O2k-MultiSensor Modules.
OroboPOS-ConnectorOroboPOS-Connector (blue POM), with male connection to OroboPOS sensor head (POS) and with cable and male plug fitting into O2k-Main Unit. Two units of this item are standard components of the O2k-Assembly Kit (O2k-FluoRespirometer), mounted on the OroboPOS-Holders at the O2k-Main Unit.
Oroboros O2k-Core (O2k-Series D - G)Oroboros O2k-Core (O2k-Series D - G) - the experimental system complete for basic high-resolution respirometry (HRR). The O2k-Core includes the O2k-Main Unit with stainless steel housing, O2k-Assembly Kit, two OroboPOS (polarographic oxygen sensors) and OroboPOS-Service Kit, DatLab software, the ISS-Integrated Suction System and the O2k-Titration Set. The O2k-Core supports all add-on O2k-Modules of the O2k.

On-line display of oxygen flux (rate of respiration) is provided in addition to the conventional 'oxygraphic' plot of oxygen concentration over time. Highest signal stability minimizes the required amounts of biological sample, and provides the basis for resolution in the extreme low-oxygen range. Peltier temperature control provides a thermal stability at ±0.002 °C in the range of 4 °C to 47 °C at typical constant room temperature. Electronically controlled PVDF or PEEK stirrers are integrated in the two-chamber design of the O2k, and a barometric pressure transducer enables automatic oxygen calibrations implemented in the DatLab software.

The O2k is a sole source apparatus with no other instruments meeting its specifications.
Oxygen sensor testPOS testThe O2 sensor test is an important component of the MitoFit Quality Control System. The OroboPOS sensor test is described in detail in MiPNet06.03 POS-calibration-SOP, is performed after switching on the Oroboros O2k, and is required as a basis of technical service of the instrument.
PC requirementsThe PC requirements for controlling an O2k and data recording with DatLab are found here.
Polarization voltageUA polarization voltage of 600 mV to 800 mV is applied between anode and cathode of the polarographic oxygen sensor, resulting in a current when oxygen is consumed. The current is converted by the electronics to a voltage (raw signal) which must not be confused with the polarization voltage.
Polarographic oxygen sensorPOSPolarographic oxygen sensors (POS) are operated with a polarization voltage between the cathode and anode, connected by an electrolyte. Cathode, anode and electrolyte are separated from the analyte by an oxygen-permeable membrane. Oxygen is reduced at the cathode such that the local oxygen concentration is maintained at zero, and diffuses along the concentration gradient from the stirred medium to the cathode, resulting in a linear calibration between oxygen partial pressure and electric current [Amp] (amperometric mode of operation). The OroboPOS is the POS applied in the Oroboros O2k.
PotentiometryPotentiometry is the general term given to the method of measuring the electric potential difference between two electrodes connected by an electrolytic solution. The potential of the reference electrode is constant. The other electrode is called the indicator electrode. If this is an ion-selective electrode which is in equilibrium with the solution, the measured electric potential difference is proportional to the (negative) logarithm of the activity of a specific ion in the solution. Examples are the pH glass electrode for measurement of pH, and the TPP+ electrode for measurement of pTPP and calculation of mt-membrane potential.
Power O2k-FluoRespirometerPower O2k-FluoRespirometer - optional configuration as additional system for increasing output combined with the O2k-FluoRespirometer (O2k-Series H). The Power O2k-FluoRespirometer includes the TIP2k and the O2k-sV-Module, and supports all add-on O2k-Modules of the Oroboros O2k. It can be added to an existing Oroboros O2k of any O2k-Series. This application does not require an additional ISS-Integrated Suction System and O2k-Titration Set. Furthermore, the OroboPOS-Mounting Tool of the OroboPOS Service Tools can be used from the available O2k and is not included.
Power O2k-Respirometer
Power-O2k

The Power O2k-Respirometer is an economical option for using additional O2k-Units to increase output in high-resolution respirometry. This application does not require an additional ISS-Integrated Suction System, an O2k-Titration Set and the O2k-Manual. Furthermore, the OroboPOS-Mounting Tool of the OroboPOS Service Tools can be used from the available O2k and is not included.

The Power O2k-Respirometer supports all add-on O2k-Modules of the Oroboros O2k.
Search for defective O2k componentsThe 2-chamber design of the O2k helps to search for defective O2k components, by switching components linked to O2k chambers A and B between sides A and B.
Shipping an O2kFor shipping an O2k, standard operating procedures have to be followed to avoid damage of the instrument and unexpected delays. The O2k-Main Unit must be shipped only in Packing\O2k-Box 1, without O2k-Chambers and without OroboPOS. Two O2k-Chamber Holders, two OroboPOS-Holders and two OroboPOS-Connectors are attached to the O2k-Main Unit for transport.
Startup O2k-RespirometerStartup O2k-Respirometer - the experimental system complete for basic high-resolution respirometry (HRR). The O2k-Respirometer includes the O2k-Main Unit with stainless steel housing, O2k-Assembly Kit, two OroboPOS (polarographic oxygen sensors) and OroboPOS-Service Kit, DatLab software, the ISS-Integrated Suction System, the O2k-Titration Set, and for performing high-resolution respirometry with reduced amounts of biological sample the O2k-sV-Module.

The O2k-Respirometer supports all add-on O2k-Modules of the Oroboros O2k, and is a component of the O2k-FluoRespirometer.

On-line display of oxygen flux (rate of respiration) is provided in addition to the conventional 'oxygraphic' plot of oxygen concentration over time. Highest signal stability minimizes the required amounts of biological sample, and provides the basis for resolution in the extreme low-oxygen range. Peltier temperature control provides a thermal stability at ±0.002 °C in the range of 4 °C to 47 °C at typical constant room temperature. Electronically controlled PVDF or PEEK stirrers are integrated in the two-chamber design of the O2k, and a barometric pressure transducer enables automatic oxygen calibrations implemented in the DatLab software.

The O2k is a sole source apparatus with no other instruments meeting its specifications.
TIP2k-ModuleTIP2k-Module - Titration-Injection microPump (TIP2k) for two-channel operation with the O2k-FluoRespirometer with automatic control by DatLab of programmable titration regimes and feedback control (oxystat, pH-stat). The TIP2k-Module includes TIP2k-Accessories: 4 Microinjection syringes, 1 Pkg. TIP2k-Needle Spacers with mounting tool, 1 TIP2k-Needle Safety Support and 1 Pkg. TIP2k Filter Papers, and the newest DatLab version.
Uninterrupted power supplyUPSA back-up power supply may be required to secure uninterrupted power supply.
Vitonn.a.Viton® is a fluoroelastomer with excellent resistance to aggressive fuels and chemicals. Viton is resistant against oxygen diffusion which makes it an ideal material for high-resolution respirometry (Viton O-rings).