MitoPedia O2k and high-resolution respirometry: O2k-Open Support
- MitoPedia: O2k-FluoRespirometry is a component of O2k-Open Support.
- It lists specific fluorometry-related topics.
- See also
|Cell count and normalization in HRR
|The cell count Nce is the number of cells, expressed in the abstract unit [x] (1 Mx = 106 x). The elementary entity cell Uce [x] is the real unit, the 'single individual cell'. A cell count is the multitude or number N of cells, Nce = N·Uce (Gnaiger MitoFit Preprints 2020.4). Normalization of respiratory rate by cell count yields oxygen flow IO2 expressed in units [amol·s-1·x-1] (=10-18 mol·s-1·x-1).
|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.
|high-resolution respirometry with the O2k.
The newest DatLab version is DatLab 8, included in the O2k-Packages. NextGen-O2k and O2k-Series J* and higher come with DatLab 8 installed on the integrated PC (Linux). DatLab 8 is required for the NextGen-O2k and is compatible with O2k-Series (H and higher).
The DatLab software is designed for 64-bit versions of Windows operating systems and does not run on MAC devices.
The minimum computer requirements are Intel-Core-2 or equivalent CPU, 2GB RAM and Windows XP (64-bit version). However, we recommend Intel i5 or equivalent CPU, 4GB RAM, Windows 10 (64-bit version) and SSD. For the proper display of DatLab on your computer, please make sure the “Language settings” are set to English.*Optionally available without integrated PC.
|Filter Set AmR
|Amplex UltraRed. These filters should be used together with Fluorescence-Sensor Green. The filter set consists of 6 LED filters (round) and 6 photodiode filters (rectangular).
|Filter Set MgG / CaG
|Magnesium green and Calcium green, respectively. These filters should be used together with Fluorescence-Sensor Blue or Smart Fluo-Sensor Blue. The filter set consists of 6 LED filters (round) and 6 photodiode filters (rectangular).
|Filter Set Saf
|Safranin. These filters should be used together with Fluorescence-Sensor Blue or Smart Fluo-Sensor Blue. The filter set consists of 6 LED filters (round) and 6 photodiode filters (rectangular).
|Fluorescence-Sensor Green and Fluorescence-Sensor Blue) and O2k-FluoRespirometer (O2k-Series H to I) 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.
|Filter-Cap equipped with Filter Set Saf for measurement of mitochondrial membrane potential with Safranin when delivered. The filter set Filter Set MgG / CaG for Magnesium green® / Calcium green® measurements is included.
|Filter-Cap equipped with Filter Set AmR for Amplex UltraRed measurements when delivered.
|Forceps for membrane application
|OroboPOS and ISE membrane application; do not use for tissue preparation.
|Forceps\stainless Steel\angular Tip\fine
|tissue preparation, stainless steel. Two pairs are used particularly for muscle fiber separation.
|Forceps\stainless Steel\rounded Tip\sharp
|tissue preparation, stainless steel, antimagnetic. One pair is recommended for placing the tissue sample onto the microbalance and for handling in combination with Forceps\stainless Steel\straight Tip\sharp.
|Forceps\stainless Steel\straight Tip\sharp
|tissue preparation, stainless steel, antimagnetic. One pair is recommended for insertion of the sample into the O2k-chamber and for handling in combination with Forceps\stainless Steel\rounded Tip\sharp.
|mitochondrial preparations and living cells combined with MultiSensor modules. Mitochondrial function and dysfunction have gained 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 the 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 pathways 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.
|ISE Package 1 TPP or Ca
|ISE-Package for 1 TPP+ and Ca2+ electrode.
|Ion-Selective Electrode TPP+ and Ca2+.
|Ion-Selective Electrode TPP+ and Ca2+.
|ISE-Inner Glass Electrode
|ISE, with Ag/AgCl- and Pt-wire
|ISE-Membrane Mounting Tool
|Ion-Selective Electrode TPP+ and Ca2+. O2k-TPP+ ISE-Module: mounting tool included.
|Ion-Selective Electrode TPP+ and Ca2+.
|ISS-Filter and Tubing
|ISS-Integrated Suction System.
|ISS-Integrated Suction System
|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.
|ISS-Waste Bottle, component of the ISS-Integrated Suction System.
|ISS-Integrated Suction System.
|Ion-Selective Electrode TPP+ and Ca2+
|ISE with 6 mm outer diameter shaft, for Stopper\white PVDF\angular Shaft\side+6.2+2.6 mm Port. O2k-TPP+ ISE-Module: 2 ISE.
|Microbalance 120 g
|wet weight determination of permeabilized fibres.
|Microsyringe\10 mm3 51/0.13 mm
|Microsyringe\100 mm3 51/0.41 mm
|Titration-Injection microPump, 200 mm3 (µl), fixed injection needle with rounded tip, with spacers.
|Microsyringe\25 mm3 51/0.15 mm
|Microsyringe\50 mm3 51/0.15 mm
|Titration-Injection microPump, 500 mm3 (µl), fixed injection needle with rounded tip, with spacers.
|MitoKit-CII/Succinate-nv and MitoKit-CII/Malonate-nv, stimulates (Snv) or inhibits (Mnanv) mitochondrial respiration in CI-deficient human blood cells, fibroblasts and heart fibres, acting on Complex II of the electron transfer system.
|O-ring sV\Viton\9.5x1 mm
|Viton\9.5x1 mm, for PEEK Stopper sV, 2 are mounted on each PEEK Stopper sV, box of 8 as spares.
|Viton\12.5x1 mm, for PVDF or PEEK O2k-Stoppers (2-mL O2k-Chamber), box of 8 as spares.
|O-ring\Viton\18x2 mm, mounted on the O2k-Chamber Holder.
|OroboPOS sensor head. Replaces the O-ring\Viton\9x1 mm
|dithionite (Na2S2O4), for zero-oxygen calibration of the OroboPOS.
|The O2k-Accessory Box contains components of the POS-Service Kit and the O2k-Assembly Kit and is shipped with the O2k.
|The O2k-Assembly Kit is a component of the Oroboros O2k, consisting of 2 PVDF Stirrer-Bars, 2 PEEK O2k-Stoppers, OroboPOS-Connectors for O2k-series A-I and NextGen-O2k series XA (attached to the O2k-Main Unit) and cables (power supply, USB-connection). Several components of the O2k-Assembly Kit are included in the O2k-Accessory Box either for shipment or for storage.
|O2k-Barometric Pressure Transducer
|O2k-Barometric Pressure Transducer, A/D converter and digital output to DatLab for continuous recording of barometric pressure [kPa or mmHg], integrated into the air calibration of the POS (MiPNet06.03 POS-calibration-SOP). Integral component of the O2k-Main Unit. The warranty on the accuracy of the signal obtained from the O2k-Barometric Pressure Transducer expires within three years.
|O2k-chamber), with O-ring\Viton\18x2 mm and V-ring\30-35-4.5 mm. Two units of this item are standard components mounted on the O2k-Main Unit.
|tissue preparation, set of 4 pairs of stainless steel, antimagnetic forceps and a pair of scissors.
|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.
|The O2k-Fluo Smart-Module 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.
|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-Fuse Power Plug\M2.5 A\5x20 mm
|O2k-Assembly Kit (O2k-FluoRespirometer), mounted on the socket for the O2k-Main Power Cable, at the rear panel of the O2k-Main Unit.
|O2k-Main Power Cable
|O2k-Main Power Cable, for connecting the main unit to the power supply.
|O2k-Main Power Cable\120 V\US-CA
|O2k-Main Power Cable\230 V\AU-NZ
|O2k-Main Power Cable\230 V\Europe
|The O2k-Main Unit is a component of the O2k-Core. The O2k-Main Unit consists of functionally defined, integral elements, the (O2k-Main Basic, O2k-Peltier Temperature Control, two O2k-Electromagnetic Stirrer Twin-Control units, two O2k-Amperometric OroboPOS Twin-Channels, O2k-Barometric Pressure Transducer), which cannot be obtained separately.
|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).
|Syringe Storage Box with Syringe Labels, a set of two Syringe Racks with Syringe Collars, and a set of two Tube Racks.
|O2k-USB Flash Drive
|Oroboros O2k containing: DatLab, O2k-Manual, O2k-Protocols, O2k-Publications, and info on O2k-Workshops.
|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.
|O2k-Window Frame from the O2k-Main Unit, for rare cleaning purposes and for front fixation of the Fluorescence-Control Unit.
|O2k-sV-Module, 12 mm inner diameter). The optical properties of Duran® allow application of fluorometric sensors (Duran® optical properties).
The OroboPOS is a polarographic oxygen sensor (POS), with an amperometric mode of operation. The OroboPOS meets the highest quality criteria in terms of linearity, stability and sensitivity of the signal. The Clark type polarographic oxygen sensor (POS) remains the gold standard for measuring dissolved oxygen in biomedical, environmental and industrial applications over a wide dynamic oxygen range.It consists of a gold cathode, a silver/silverchloride anode and a KCl electrolyte reservoir separated from the sample by a 25 µm membrane (FEP). The main body of the OroboPOS is made of PEEK. With application of a polarization voltage (0.8 V), a current is obtained as an amperometric signal, which is converted to a voltage.
|OroboPOS head (POS) and with cable and male plug fitting into O2k-Main Unit.
|The OroboPOS-Connector Service entails routine maintenance and any necessary repairs of the OroboPOS-Connector in the Oroboros electronics workshop (WGT).
Oroboros O2k - the modular system for high-resolution respirometry (HRR) for mitochondria and cell research. Oroboros delivers the O2k-technology for high-resolution respirometry in mitochondria and cell research. The O2k allows the measurement of respiration at controlled oxygen levels, combined with redox biology (NADH and CoQ), ROS production, mitochondrial membrane potential, ATP production, Ca2+, or pH. HRR expands to HRPB: High-Resolution PhotoBiology.
Small amounts of biological samples can be used for bioenergetic and OXPHOS analysis, ranging from isolated mitochondria, permeabilized tissues and permeabilized cells to living cells and tissues slices.The modular O2k-concept is supported by DatLab, with high flexibility for extension by add-on O2k-Modules. The globally tested and trusted high-resolution O2k technology prioritizes both quality and scientific research output in the field of mitochondrial physiology and pathology.
|oxygen and hydrogen by electrolysis of water using a proton exchange membrane (PEM). O2 and H2 gas can be used to control the O2 regime in the Oroboros O2k (Setting_the_oxygen_concentration) using the 10 mL Gas-Injection Syringes. Low oxygen concentrations (<50 µM) are used to mimic tissue normoxia or hypoxia. Hyperoxic conditions above air saturation (250-600 µM O2) are routinely used for high-resolution respirometry of permeabilized muscle fibers or to induce oxidative stress in cells and mitochondrial preparations.
|tissue homogenate preparation
|PBI-Shredder O2k-Set. Select 230 V or 120 V. Oroboros Instruments: world-wide distributor.
|POS-Electrolyte Powder, KCl. The powder is dissolved in 10 ml distilled water to yield a 1 M KCl solution.
|O2k-Main Unit, guiding theguiding the POS to the 2-mL O2k-chamber, and keeping the SmartPOS/OroboPOS-Connector in a fixed position for sealing the O2k-chamber with the POS-Seal Tip. In addition, the POS-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.
|POS-Membrane Ring, PEEK, holds the membrane against the inner O-ring on the POS housing.
|POS-Membranes, FEP 25 µm; 40/Pck.
|POS-Mounting Tool for application of POS-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 POS-Membrane Ring during membrane application. Since the O2k-Series G-0075 membrane ring holder have a 2 mm extended ridge for a better grip.
|POS (sensor head) against the O2k-Chamber. Push the wetted POS-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.
|POS-Service Kit, in O2k-Accessory Box including all oxygen sensor service accessories for membrane mounting and service of the POS.
|O2k-Main Unit, with polystyrene inlets. Keep the original packing material safely stored, for any future shipping purposes of the O2k-Main Unit.
|Packing\O2k-Box 1+2 for shipping the O2k-Core. O2k-WorldWide delivery, insurance and handling are included in the O2k-Core.
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
|Microsyringe\10 mm3 51/0.13 mm, spare.
|Power O2k Numbers
|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.
|Q redox state and cyclic voltammetry, is supported by the NextGen-O2k and consists of the Q-Sensor, integrated electronic components in the O2k, and the DatLab software.
|Reference-Electrode\2.4 mm, 2.4 mm diameter glass.
|Scissors\stainless steel\straight Tip\sharp
|Screwdriver allen wrench
|O2k-FluoRespirometer and O2k-sV-Module.
|Shredder-Tube Ram Tool
|Shredder-Tube Ram Tool and Shredder-Tube Cap Tool, coral colour (Box of 100) and Shredder Tube FT500-PMS with Lysis Disk, serrated Shredder-Tube Ram Tool and Shredder-Tube Cap Tool, coral colour (Box of 100). There is no difference between the white and the coral shredder tubes, except that the white tubes are also approved for high pressure.
|Smart Fluo-Sensor Blue
|Filter-Cap equipped with Filter Set Saf for measurement of mitochondrial membrane potential with safranin and rhodamine 123. Filter Set MgG / CaG for Magnesium green and Calcium green measurements are included.
|Smart Fluo-Sensor Green
|Filter-Cap equipped with Filter Set AmR for Amplex UltraRed and TMRM measurements when delivered.
The SmartPOS is a polarographic oxygen sensor (POS), for O2k-Series J and XB onward, with an amperometric mode of operation. The SmartPOS combines the previous OroboPOS and OroboPOS-Connector and is automatically recognized by the software DatLab 8. Combination of the previously separated components in one piece, protects the electrical connections. The SmartPOS is, like the OroboPOS, a Clark type polarographic oxygen sensor (POS), which remains the gold standard for measuring dissolved oxygen in biomedical, environmental and industrial applications over a wide dynamic oxygen range. The SmartPOS meets the highest quality criteria in terms of linearity, stability and sensitivity of the signal.The sensor consists of a gold cathode, a silver/silverchloride anode and a KCl electrolyte reservoir separated from the sample by a 25 µm membrane (FEP). With application of a polarization voltage (0.8 V), a current is obtained as an amperometric signal.
|Stirrer-Bar sV\white PVDF\11.5x6.2 mm
|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
|PEEK\15x6 mm, for 2-mL O2k-Chamber, for specific optical applications to minimize optical interference.
|Stirrer-Bar\white PVDF\15x6 mm
|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
|Volume-Calibration Ring sV (A or B) for volume adjustment 0.5 mL; 2 mounted O-rings (O-ring sV\Viton\9.5x1 mm).
|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
|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
|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.
|Stopper\black PEEK\conical Shaft\central Port\peripheral 2.3 mm Port
|O2k-MultiSensor applications (NO, H2S, H2O2); 1 additional port (2.3 mm); central titration port; conical 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+2.3+2.6 mm Port
|Syringe Storage Box
|O2k-chamber. 2 syringes are supplied with Oxia.
|Syringe\500 mm3 51/0.41 mm
|TIP2k Filter Papers
|O2k-FluoRespirometer with automatic control by DatLab of programmable titration regimes and feedback control (oxystat, pH-stat).
|TIP2k-Needle Safety Support with cable guide
|DatLab of programmable titration regimes and feedback control (oxystat, pH-stat).
|O2k-chambers; package of two (for a total of eight tubes).
|USB-Cable 2.0\Type A-B
|O2k-Main Unit (O2k-Series E upwards) to the USB port of a PC or laptop.
|O2k-Chamber Holder and O2k-Chamber Holder sV.
|O2k-chamber), white; attached to the stoppers.
|Volume-Calibration Ring sV
|PEEK O2k-Stopper sV.