MitoPedia: O2k-Open Support

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OROBOROS support
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MitoPedia: O2k-Open Support

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O2k-Open Support

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

MitoPedia: O2k-Open Support list Frequently Asked Questions and keywords for help.
More details - »MitoPedia: O2k hardware« - »MitoPedia: DatLab« - »O2k-Open Support alert«

ATPTAdenosine triphosphate is a nucleotid and functions as the major carrier of chemical energy in the cells. As it transfers its energy to other molecules, it looses its terminal phosphate group and becomes adenosine diphosphate (ADP).
Amplex UltraRedAmRAmplex UltraRed (AmR) is used as an extrinsic fluorophore for measurement of hydrogen peroxide production (ROS) by cells or mitochondrial preparations. The reaction of H2O2 and AmR is catalyzed by horseradish peroxidase to produce the red fluorescent compound resorufin (excitation wavelength 563 nm, emission 587 nm; the fluorescenet product according to the supplier is called UltroxRed in the case of Amplex UltraRed which has a very similar structure to resorufin). The change of emitted fluorescence intensity is directly proportional to the concentration of H2O2 added, whereby the H2O2 is consumed.
Attached cellsMany cell types are grown in culter as attached cells, such as endothelial or neuronal cells in a monolayer.
Biological contaminationBiological contamination may be caused by microbial growth in the O2k-Chamber or in the experimental medium.
CalciumCaCa2+ is a major signaling molecule in both prokaryotes and eukaryotes. Its cytoplasmic concentration is tightly regulated by transporters in the plasma membrane and in the membranes of various organelles. For this purpose it is either extruded from the cell through exchangers and pumps or stored in organelles such as the endoplasmic reticulum and the mitochondria. Changes in the concentration of the cation regulate numerous enzymes including many involved in ATP utilizing and in ATP generating pathways and thus ultimately control metabolic activity of mitochondria and the of entire cell. Measuring changes in Ca2+ levels is thus of considerable interest in the context of high-resolution respirometry.
Calcium GreenCaGCalcium Green denotes a family of extrinsic fluorophores applied for measurement of Ca2+ concentration.
Closed chamberCThe O2k chamber can be used as a closed system or open system. Gas bubbles must be avoided.
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A Cover-Slip should be placed on top of the O2k-Stopper to minimize contamination and evaporation of liquid extruding from the capillary of the stopper. The Cover-Slips do not exert any direct effect on oxygen backdiffusion into the O2k-chamber. Use the the Cover-Slip\black to avoid light penetration and disturbance of fluorescence signals and generally for optical measurements in the O2k.
DatLab 2DL2DatLab 2 (DL2) is a MS-DOS programe. DL2 is still used for analysis of oxygen kinetics, after exporting files recorded in recent DatLab versions. A user-friendly O2-kinetics module is in preparation (DL8).
DatLab oxygen flux: performance and data analysisThe quality of the results are strongly affected by the performance and data analysis. Therefore, we provide guidelines for performing and evaluating respirometric assays.
Different O2 fluxes in left and right chamberWhat are potential causes for different O2 fluxes in the left and right chamber?
DithioniteDithDithionite Na2S2O4 is the 'zero oxygen solution powder' used for calibration of oxygen sensors at zero oxygen, or for stepwise reduction of oxygen concentrations in instrumental O2 background tests. It is not recommended to use dithionite in experiments with biological samples or several multisensor approaches, for these see Setting the oxygen concentration.
Enable DL-Protocol editingIn DatLab 7.4, it is possible to edit a DL-Protocol and save it as DLPU. To enable it, select in the menu 'Protocols' the option 'Enable DL-Protocol editing', then select the plot in which the marks will be set (e.g., O2 flux per V). Select the 'Overview' window, where you will be able to edit events and marks names, definition/state, final concentration and titration volumes, as well as select a mark as 'multi' for multiple titration steps, skip a mark, or add a new event or mark. After saving, export a DL-Protocol User (DLPU) and load it before running the next experiments.

For more information:

PlayVideo.jpg Export DL-Protocol User (*.DLPU)
Export DL-Protocol User (*.DLPU)Export DL-Protocol User (*.DLPU) Protocol possess unique D### codes and comprise a fixed sequence of events and marks which cannot be changed by the user. However, the user may edit concentrations and titration volumes of injections and store the modified protocol as user-specific DL-Protocol [File]\Export\DL-Protocol User (*.DLPU).

If users of older DatLab versions than DatLab 7.4 wish to alter the nature of the chemicals used or the sequence of injections, we ask them to contact the O2k-Technical Support.

DatLab 7.4

DatLab 7.4 offers a new feature in DL-Protocols: flexibility. Fixed sequence of events and marks can be changed (Skip/Added) in a SUIT protocol by the user. Moreover, editions of text, instructions, concentrations and titration volumes of injections in a specific DL-Protocol can be edited and saved as user-specific DL-Protocol [File]\Export\DL-Protocol User (*.DLPU).

For more information, see: Enable DL-Protocol editing
Fatty acid oxidationFAO'Fatty acid oxidation (β-oxidation) is a multi-step process by which fatty acids are broken down to generate acetyl-CoA, NADH and FADH2 for further energy transformation. Fatty acids (short chain with 4–8, medium-chain with 6–12, long chain with 14-22 carbon atoms) are activated by fatty acyl-CoA synthases (thiokinases) in the cytosol. The mt-outer membrane enzyme carnitine palmitoyltransferase I (CPT 1) generates an acyl-carnitine intermediate for transport into the mt-matrix. Octanoate, but not palmitate, (eight- and 16-carbon saturated fatty acids) may pass the mt-membranes, but both are frequently supplied to mt-preparations in the activated form of octanoylcarnitine or palmitoylcarnitine. Electron-transferring flavoprotein complex (CETF) is located on the matrix face of the mt-inner membrane, and supplies electrons from fatty acid β-oxidation (FAO) to CoQ.
Filter-CapFilter-Cap: O2k-Fluo LED2-Module (O2k-Series D to G) sensors (Fluorescence-Sensor Green and Fluorescence-Sensor Blue) and O2k-FluoRespirometer (O2k-Series H) sensors (Smart Fluo-Sensor Green and Smart Fluo-Sensor Blue) are equipped with a removable Filter-Cap for exchange of optical filters for the optical pathways from the LED to the sample and from the sample to the photodiode.
Fluorescence-Control UnitFluorescence-Control Unit with O2k-Front Fixation, Current-Control (O2k-Chamber A and B) for regulation of light intensity of the LED in the fluorescence sensors. This item is a standard component of the O2k-Fluorescence LED2-Module.
Forceps for membrane application
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Forceps for membrane application: for OroboPOS and ISE membrane application; do not use for tissue preparation.
GainThe gain is an amplification factor applied to an input signal to increase the output signal.
High signal at zero oxygenR0'A high signal at zero oxygen may be observed during zero calibration. The following instructions show how to further localize and possibly solve this technical problem.
Hydrogen peroxideH2O2
Hydrogen peroxide
Hydrogen peroxide, H2O2 or dihydrogen dioxide, is one of several reactive oxygen intermediates generally referred to as reactive oxygen species (ROS). It is formed in various enzyme-catalyzed reactions (e.g., superoxide dismutase) with the potential to damage cellular molecules and structures. H2O2 is dismutated by catalase to water and oxygen. H2O2 is produced as a signaling molecule in aerobic metabolism and passes membranes more easily compared to other ROS.
ISE Package 1 TPP or Ca
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O2k-TPP+ and Ca2+ ISE\1 Chamber: ISE-Package for 1 TPP+ and Ca2+ electrode.
ISS-Filter and Tubing
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ISS-Filter and Tubing, ISS-Integrated Suction System.
ISS-Integrated Suction System
ISS-Integrated Suction System: Suction pump 230 or 120 V with stainless steel housing, 2 liter waste bottle, filter and tubing; for siphoning off excess medium from the O2k-Stopper and for emptying the O2k-chambers. The ISS is included as a standard component of the O2k-FluoRespirometer. Media containing living cells or microorganisms, various poisons (inhibitors, uncouplers) and mixtures of proteins and substrates are safely disposed off in the 2-litre waste bottle. For specific country configurations, see Product ID 28101, 28102, 28103.
ISS-Lid, for ISS-Waste Bottle, component of the ISS-Integrated Suction System.
ISS-Waste Bottle
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ISS-Waste Bottle, 2-liter, component of the ISS-Integrated Suction System.
IlluminationF10The chambers of the Oroboros O2k are illuminated by an internal LED. The illumination is switched on and off in DatLab during the experiment by pressing [F10]. This illumination must be distinguished from light introduced into the chambers by LEDs for the purpose of spectrophotometric and fluorometric measurements. For these, the internal illumination must be switched off.
Living cellsvceCell viability in living cells should be >95 % for various experimental investigations, including cell respirometry. Viable cells (vce) are characterized by an intact plasma membrane barrier function. The total cell count (Nce) is the sum of viable cells (Nvce) and dead cells (Ndce). In contrast, the plasma membrane can be permeabilized selectively by mild detergents (digitonin), to obtain the mt-preparation of permeabilized cells used for cell ergometry. Living cells are frequently labelled as intact cells in the sense of the total cell count, but intact may suggest dual meanings of viable or unaffected by a disease or mitochondrial injury.
Magnesium GreenMgGMagnesium Green (MgG) belongs to the extrinsic fluorophores applied for measurement of mitochondrial ATP production with mitochondrial preparations. This dye fluoresces when bound to Mg2+. The technique to measure mitochondrial ATP production is based on the fact that Mg2+ presents different dissociation constants for ADP and ATP, and the adenine nucleotide translocase (ANT) exchanges ATP for ADP.
Mitochondrial Respiration Medium - MiR05-Kit, 1 vial; for a final volume of 250 mL
Microbalance 120 g
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Microbalance 41 g to 120 g; 0.01 mg / 0.1 mg display; particularly for wet weight determination of permeabilized fibres.
Microsyringe\200 mm3\TIP2k
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Microsyringe\200 mm3\TIP2k: Microinjection syringe for Titration-Injection microPump, 200 mm3 (µl), fixed injection needle with rounded tip, with spacers.
Microsyringe\500 mm3\TIP2k
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Microsyringe\500 mm3\TIP2k: Microinjection syringe for Titration-Injection microPump, 500 mm3 (µl), fixed injection needle with rounded tip, with spacers.
Cell permeable compound - MitoKit-CII, 2 vials; 1 vial of plasma membrane-permeable succinate-nv (Snv; NV118; 25 mg) and 1 vial of plasma membrane-permeable malonate-nv (Mnanv; NV161; 10 mg).
MitoPedia: DatLab
MitoPedia: O2k hardware
MitoPedia: O2k-FluoRespirometry
MitoPedia: O2k-Open Support
MitoPedia: O2k-Respirometry
MitoPedia: SUIT
Mitochondrial membrane potentialmtMP, Δψ [V]The mitochondrial membrane potential, mtMP, is the electric part of the protonmotive force, ΔpH+.

Δψ = ΔpH+ - ΔµH+ / F

mtMP or Δψ is the potential difference across the inner mitochondrial (mt) membrane, expressed in the electric unit of volt [V]. Electric force of the mitochondrial membrane potential is the electric energy change per ‘motive’ electron or per electron moved across the transmembrane potential difference, with the number of ‘motive’ electrons expressed in the unit coulomb [C].
Motic Microscope
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Motic Microscope SMZ-171 TP: for preparation of permeabilized fibres; compact and light stereo microscope, Greenough optical system, switching power supply for use worldwide (100-240V); including auxiliary ESD objective 2.0X(38.6mm).
O-ring sV\Viton\9.5x1 mm
O-ring sV\Viton\9.5x1 mm, for PEEK Stopper sV, 2 are mounted on each PEEK Stopper sV, box of 8 as spares.
O-ring\Viton\12.5x1 mm
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O-ring\Viton\12.5x1 mm, for PVDF or PEEK O2k-Stoppers (2-mL O2k-Chamber), box of 8 as spares.
O2-Zero Powder
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O2-Zero Powder, dithionite (Na2S2O4), for zero-oxygen calibration of the OroboPOS.
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-Chamber sV
O2k-Chamber sV: 12 mm inner diameter, Duran® glass polished, with standard operation volume V of 0.5 mL.
O2k-Dissection Set
O2k-Dissection Set: for tissue preparation, set of 4 pairs of stainless steel, antimagnetic forceps and a pair of scissors.
O2k-Fluo LED2-Module
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The 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, Rhodamine 123), Ca2+ (Calcium Green™), and numerous other applications open for O2k-user innovation.
O2k-Fluo Smart-Module
The 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.
The 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.
O2k-NO Amp-Module
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O2k-NO Amp-Module: NO-sensor compatability pack an amperometric add-on for O2k-MultiSensor application The NO sensor is not included.
O2k-Open Support
O2k-Open Support agreementO2k-Open Support aims at providing expert help quickly. Please, help us sharing our support communication openly with the scientific community.
O2k-OroboPOS-Holder MitoPediaThe OroboPOS-Holder (POS-Holder), made from blue POM, is screwed into the copper block of the O2k-Main Unit, guiding the OroboPOS-Connector with the OroboPOS sensor head (POS) to the O2k-Chamber, and keeping the OroboPOS-Connector in a fixed position for sealing the O2k-Chamber with the OroboPOS-Seal Tip. In addition, the OroboPOS-Holder fixes the O2k-Chamber in an accurate rotational position by pressing against the angular cut of the glass chamber. Two units of this item are standard components mounted on the O2k-Main Unit.
O2k-Peltier MitopediaO2k-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-TPP+ ISE-Module
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O2k-TPP+ ISE-Module: Potentiometric ion-selective electrodes for measurement of mitochondrial membrane potential
O2k-Titration Set
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The O2k-Titration Set consists of Hamilton microsyringes (6 x 10 mm3 and 3 spare plungers, 6 x 25 mm3, 1 x 50 mm3, 1 x 100 mm3, 1 x 500 mm3; fixed needles with rounded tips), provided in the Syringe Storage Box with Syringe Labels, a set of two Syringe Racks with Syringe Collars, and a set of two Tube Racks. The O2k-Titration Set is included in the O2k-FluoRespirometer and Startup O2k-Respirometer.
O2k-Window FrameO2k-Window Frame: blue POM, with thread for fixation on the O2k-Main Unit, to be removed only for rare cleaning purposes and for front fixation of the Fluorescence-Control Unit, using the O2k-Window Tool. Two units of this item are standard components of the (Oroboros O2k-Core (O2k-Series D - G) ), mounted on the O2k-Main Unit.
O2k-Chamber: Duran® glass polished, with standard operation volumes (V) of 2.0 mL or 0.5 mL (small chamber volume in the O2k-sV-Module, 12 mm inner diameter). The optical properties of Duran® allow application of fluorometric sensors (Duran® optical properties).
O2k-pH ISE-Module
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O2k-pH ISE-Module: two pH electrodes and reference electrodes and accessories
The O2k-sV-Module is the O2k small-volume module, comprised of two Duran® glass chambers of 12 mm inner diameter specifically developed to perform high-resolution respirometry with reduced amounts of biological sample, and all the components necessary for a smaller operation volume V of 0.5 mL. The current DatLab version is included in the delivery of this revolutionary module.
Open chamberOThe term "open O2k-chamber" refers to a situation in which the liquid phase is allowed to equilibrate with a gas phase, but the stopper is partially inserted using the Stopper-Spacer.
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OroboPOS-Connector (blue POM), with male connection to OroboPOS sensor head (POS) and with cable and male plug fitting into O2k-Main Unit.
OroboPOS-Connector MitoPediaOroboPOS-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.
OroboPOS-Electrolyte Powder
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OroboPOS-Electrolyte Powder, KCl. The powder is dissolved in 10 ml distilled water to yield a 1 M KCl solution.
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The OroboPOS-Holder (POS-Holder), made from blue POM, is screwed into the copper block of the O2k-Main Unit, guiding the OroboPOS-Connector with the OroboPOS sensor head (POS) to the 2-mL O2k-chamber, and keeping the OroboPOS-Connector in a fixed position for sealing the O2k-chamber with the OroboPOS-Seal Tip. In addition, the OroboPOS-Holder fixes the O2k-Chamber in an accurate rotational position by pressing against the angular cut of the glass chamber. Two units of this item are standard components mounted on the O2k-Main Unit.
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OroboPOS-Membranes, FEP 25 µm; 40/Pck.
OroboPOS-Mounting Tool
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OroboPOS-Mounting Tool for application of OroboPOS-Membranes. It consists of two parts, (i) the membrane guide (larger diameter) and (ii) the membrane ring holder with O-ring\Viton\6x1 mm for holding the OroboPOS-Membrane Ring during membrane application. Since the O2k-Series G-0075 membrane ring holder have a 2 mm extended ridge for a better grip.
OroboPOS-Polishing Cloth
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OroboPOS-Polishing Cloth for cathode cleaning. Replace the OroboPOS-Polishing Cloth at intervals. A two-year interval may be considered in cases of intensive use.
OroboPOS-Polishing Powder
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OroboPOS-Polishing Powder for cathode cleaning.
OroboPOS-Seal Tip
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OroboPOS-Seal Tip, black butyl rubber gasket with 3 mm pore. 4/Pck, for sealing the OroboPOS (sensor head) against the O2k-Chamber. Push the wetted OroboPOS-Seal Tip over the POS-sensor head, with the pore positioned centrally, not covering any peripheral area of the gold cathode. Do not stretch the gasket across the POS-sensor head.
OroboPOS-Service Kit
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OroboPOS-Service_Kit, with O2k-Service Box including all oxygen sensor service accessories for membrane mounting and service of the OroboPOS.
Oxygen calibration - DatLabO2 calibration is the calibration in DatLab of the oxygen sensor. It is a prerequisite for obtaining accurate measurements of respiration. Accurate calibration of the oxygen sensor depends on (1) equilibration of the incubation medium with air oxygen partial pressure at the temperature defined by the experimenter; (2) zero oxygen calibration; (3) high stability of the POS signal tested for sufficiently long periods of time; (4) linearity of signal output with oxygen pressure in the range between oxygen saturation and zero oxygen pressure; and (5) accurate oxygen solubility for aqueous solutions for the conversion of partial oxygen pressure into oxygen concentration. The standard oxygen calibration procedure is described below for high-resolution respirometry with the automatic calibration routine using instrumental calibraiton DL-Protocols in DatLab.
Oxygen flux - instrumental backgroundJ°O2Instrumental background oxygen flux, J°O2, in a respirometer is due to oxygen consumption by the POS, and oxygen diffusion into or out of the aqueous medium in the O2k-chamber. It is a property of the instrumental system, measured in the range of experimental oxygen levels by a standardized instrumental O2 background test. The oxygen regime from air saturation towards zero oxygen is applied generally in experiments with isolated mitochondria and living or permeabilized cells. To overcome oxygen diffusion limitation in permeabilized fibers and homogenates, an elevated oxygen regime is applied, requiring instrumental background test in the same range of elevated oxygen.
Oxygen kineticsOxygen kinetics describes the dependence of respiration of isolated mitochondria or cells on oxygen partial pressure. Frequently, a strictly hyperbolic kinetics is observed, with two parameters, the oxygen pressure at half-maximum flux, p50, and maximum flux, Jmax. The p50 is in the range of 0.2 to 0.8 kPa for cytochrome c oxidase, isolated mitochondria and small cells, strongly dependent on Jmax and coupling state.
Oxygen sensor testPOS testThe O2 sensor test is an important component of the MitoFit Quality Control System. The OroboPOS 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.
Oxygen signalThe oxygen signal of the Oroboros O2k is transmitted from the electrochemical polarographic oxygen sensor (OroboPOS) for each of the two O2k-chambers to DatLab. The primary signal is a current [µA] which is converted into a voltage [V] (raw signal), and calibrated in SI units for amount of substrance concentration [µmol·L-1 or µM].
PBI-Shredder MitoPediaPBI-Shredder: Auxiliary O2k-Tool for tissue homogenate preparation with high mitochondrial yield and function.
PBI-Shredder O2k-Set
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PBI-Shredder O2k-Set: Auxiliary O2k-Tool for tissue homogenate preparation; the Shredder-Kit Box contains the heavy duty high torque SG3 driver with convertible handle, SG3 base with 3 position force setting lever (FSL), battery charger, two lithium ion batteries, Shredder-Tube Cap Tool and Shredder-Tube Ram Tool. The PBI-Shredder O2k-Set includes the Shredder-Kit Box with 100 Shredder-Tubes FT500-PS, 100 Shredder-Tubes FT500-PMS, a pair of sharp forceps for tissue dissection, a pair of scissors and a manual.
PH calibration bufferspH calibration buffers are prepared to obtain two or more defined pH values for calibration of pH electrodes and pH indicator dyes.
Packing\Peli Case
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Packing\Peli Case: Watertight, crushproof, and dust proof case for safe transportation of the O2k:

Retractable extension handle, Strong polyurethane wheels with stainless steel bearings, Easy open Double Throw latches, Open cell core with solid wall design - strong, light weight, O-ring seal, Automatic Pressure Equalization Valve, Fold down handles, Stainless steel hardware and padlock protectors, 4 level Pick N Pluck™ with convoluted lid foam, Special foam inlets for O2k.

Interior Dimensions: 21.48" x 16.42" x 12.54" (54.5 x 41.7 x 31.8 cm)
Pipette\Plastic\1 ml ungraded
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Pipette\Plastic\1 mL ungraded, for filling electrolyte into the reservoir of the OroboPOS.
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.
Power O2k Numbers
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Power O2k Numbers: Single number to label the O2k in a Power O2k-Lab.
Power O2k-FluoRespirometer
Power O2k-FluoRespirometer
Power 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
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The Power O2k-Respirometer is an economical option for using additional O2k-Units to increase output in high-resolution respirometry.
Proton fluxJH+Volume-specific proton flux is measured in a closed system as the time derivative of proton concentration, expressed in units [pmol·s-1·mL-1]. Proton flux can be measured in an open system at steady state, when any acidification of the medium is compensated by external supply of an equivalent amount of base. The extracellular acidification rate (ECAR) is the change of pH in the incubation medium over time, which is zero at steady state. Volume-specific proton flux is comparable to volume-specific oxygen flux [pmol·s-1·mL-1], which is the (negative) time derivative of oxygen concentration measured in a closed system, corrected for instrumental and chemical background. pH is the negative logarithm of proton activity. Therefore, ECAR is of interest in relation to acidification issues in the incubation buffer or culture medium. The physiologically relevant metabolic proton flux, however, must not be confused with ECAR.
Raw signal of the oxygen sensorRThe raw signal of the polarographic oxygen sensor is the voltage obtained after a current-to-voltage conversion in the O2k. O2k-Support page: Oxygen signal
SUITbrowserUse the SUITbrowser to find the best substrate-uncoupler-inhibitor-titration (SUIT) protocol for your research questions. Open the SUITbrowser:
SafraninSafSafranin is one of the most established dyes for measuring mitochondrial membrane potential by fluorometry. It is an extrinsic fluorophore with an excitation wavelength of 495 nm and emission wavelength of 587 nm. Safranin is a potent inhibitor of N-linked respiration and of the phosphorylation system. Synonyms: Safranin O, Safranin Y, Safranin T, Gossypimine, Cotton Red, Basic Red2
Screwdriver allen wrench
Screwdriver allen wrench, a standard component of the O2k-FluoRespirometer and O2k-sV-Module.
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.
Setting the oxygen concentration
Shipping an O2kFor shipping an O2k or parts, 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-Respirometer
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Startup 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 is a sole source apparatus with no other instruments meeting its specifications.
Stirrer-Bar sV\white PVDF\11.5x6.2 mm
Stirrer-Bar sV\white PVDF\11.5x6.2 mm, operated in the 0.5-mL O2k-Chamber sV at constant stirring speed (standard is 750 rpm, or 12.5 Hz), to provide optimum mixing of the sample in the aqueous medium and ensure a stable signal of the polarographic oxygen sensor (OroboPOS) placed in a position of maximum current of the medium.
Stirrer-Bar\black PEEK\15x6 mm
Stirrer-Bar black PEEK15x6 mm.JPG
Stirrer-Bar\black PEEK\15x6 mm, for 2-mL O2k-Chamber, for specific optical applications to minimize optical interference.
Stirrer-Bar\white PVDF\15x6 mm
Stirrer bar PVDF.jpg
Stirrer-Bar\white PVDF\15x6 mm, operated in the 2-mL O2k-chamber at constant stirring speed (standard is 750 rpm, or 12.5 Hz), to provide optimum mixing of the sample in the aqueous medium and ensure a stable signal of the polarographic oxygen sensor (OroboPOS) placed in a position of maximum current of the medium.
Stopper sV\black PEEK\conical Shaft\central Port
Stopper sV\black PEEK\conical Shaft\central Port: with conical shaft (with PTFE, graphite, carbon fiber) and one central capillary (1.0 mm diameter; 48.9 mm length), Volume-Calibration Ring sV (A or B) for volume adjustment 0.5 mL; 2 mounted O-rings (O-ring sV\Viton\9.5x1 mm).
Gas spacer for Stopper.jpg
Stopper-Spacer (or gas spacer) to set the O2k-Stopper into a standard position for a fixed gas phase above the aqueous phase in the 2-mL O2k-chamber, during air calibration or for injection of nitrogen, argon or oxygen into the gas phase; Stopper-Spacer thickness of 4mm (a deviation of +/- 10% possible).
Stopper\black PEEK\angular Shaft\side+6.2+2.6 mm Port
Stopper black PEEK angular Shaft side+6.2+2.6 mm Port.JPG
Stopper\black PEEK\angular Shaft\side+6.2+2.6 mm Port, for application with ISE; side titration port and two additional holes (6.2 mm and 2.6 mm); angular bottom; including Volume-Calibration Ring (A or B); 2 mounted O-rings, with 8 spare O-rings (O-ring\Viton\12.5x1 mm).
Stopper\black PEEK\conical Shaft\central Port
Stopper black PEEK conical Shaft central Port.JPG
Stopper\black PEEK\conical Shaft\central Port: with conical shaft (with PTFE, graphite, carbon fiber) and one central capillary (1.3 mm diameter; 50.6 mm length), Volume-Calibration Ring (A or B) for volume adjustment 1.5 to 3.2 ml; 2 mounted O-rings, with 8 spare O-rings (O-ring\Viton\12.5x1 mm). The black PEEK stoppers are required for optical O2k-MultiSensor Modules.
Syringe\60 ml\Gas-Injection
Gas injection syringe.jpg
Syringe\60 mL\Gas-Injection, 60 mL, with spacer and stainless steel needle, flat tip, for gas injection into the O2k-chamber.
TIP2k Filter Papers
TIP2k Filter Papers.JPG
TIP2k Filter Papers (package of 10) to be used for the stoppers during TIP2k titration for Instrumental oxygen background correction to avoid constant manual siphoning off of extruded liquid.
TIP2k syringe blockedWhen the TIP2k syringe is blocked, it must not be used with the TIP2k, and specific cleaning instructions should be followed.
TIP2k with 200 mm3 microsyringe.JPG
TIP2k-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).
TIP2k-Needle Safety Support with cable guide
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TIP2k-Needle Safety Support with cable guide: for safe storage of TIP2k-needles and MultiSensor Modules cables when not required during the experiment.
TIP2k-Needle Spacer
80600-24 Spacers f. TIP2k needle.JPG
TIP2k-Needle Spacers for microinjection TIP2k needle, silicone stops (200/Pkg.) with mounting tool.
TMRMTMRMTMRM (tetramethylrhodamine methyl ester) is an extrinsic fluorophore used as a probe to determine changes in mitochondrial membrane potential. TMRM is a lipophilic cation that is accumulated in the mitochondrial matrix in proportion to Δψmt. Upon accumulation of the dye it exhibits a red shift in its absorption and fluorescence emission spectrum. The fluorescence intensity is quenched when the dye is accumulated in the mitochondrial matrix.
Temperature plot emptyProblem: Layout "01 Calibration Exp. Gr3-Temp" is selected, a third plot is displayed on the screen but it remains empty (no plot is shown). Newer versions of DatLab include pre-installed layouts which do not recognize some channel designations from older O2k series.
TetraphenylphosphoniumTPP+Tetraphenylphosphonium (TPP+). A lipophilic molecular probe in conjunction with an ion selective electrode (ISE) for measuring the mitochondrial membrane potential.
Time resolutionTime resolution in respirometric measurements is influenced by three parameters: the response time of the POS, the data sampling interval and the number of points used for flux calculation.
Titration-Injection microPump
Titration-Injection microPump (TIP2k) for two-channel operation with the O2k with automatic control by DatLab of programmable titration regimes and feedback control (oxystat, pH-stat).
Uncoupler titrationsIn uncoupler titrations various uncouplers, such as CCCP, FCCP or DNP are applied to uncouple mitochondrial electron transfer from phosphorylation (ATP synthase, ANT and phosphate carrier), particularly with the aim to measure ET capacity. ET capacity is maximum oxygen flux measured as noncoupled respiration with optimum uncoupler concentration.
Uninterrupted power supplyUPSA back-up power supply may be required to secure uninterrupted power supply.
Virtual O2k-Workshop self-study material
Voltage (raw signal) of the oxygen sensorRThe raw signal of the polarographic oxygen sensor is the voltage obtained after a current-to-voltage conversion in the O2k. O2k-Support page: Oxygen signal
Volume-Calibration Ring
Volume calibration ring A+B PVDF for Stopper.jpg
Volume-Calibration Ring (A or B) PVDF for O2k-Stoppers (2-mL O2k-chamber), white; attached to the stoppers.
Volume-Calibration Ring sV
Volume calibration ring A+B PVDF for Stopper.jpg
Volume-Calibration Ring sV (A or B), white PVDF, attached to PEEK O2k-Stopper sV.
Zero calibrationR0Zero calibration is together with air calibration one of the two steps of the OroboPOS calibration. It is performed in the closed chamber after all the oxygen has been removed by the addition of dithionite or by respiration of imt or cells. Any incubation medium can be used for zero calibration with dithionite or sample. Unlike air calibration it is not necessary to perform a zero calibration each day.
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