| Term | Abbreviation | Description |
|---|
| Cell count and normalization in HRR | Nce | 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). |
| Chamber-Holder | | The Chamber-Holder of the Oroboros holds the Oroboros chamber in a tight vertical position with pressure exerted on the cV O-ring (18x2 mm) or sV O-ring (16x2 mm). The V-ring prevents liquid to flow from the stainless steel housing into the space outside of the chamber. |
| Cover-Slip\black | | A Cover-Slip should be placed on top of the Oroboros 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 Oroboros Chamber. Use the the Cover-Slip\black to avoid light penetration and disturbance of fluorescence signals and generally for optical measurements in the Oroboros. |
| DatLab | | DatLab is the Oroboros Software for Data Acquisition & Analysis, specifically developed for high-resolution respirometry. The Oroboros NextGen-O2k and the Oroboros O2k (O2k-Series J and higher, optionally available without integrated PC) are delivered with DatLab 8 installed on the integrated PC (Linux). DatLab 8 is included in all Oroboros Packages. The newest DatLab version is DatLab 8.2, compatible with all NextGen-O2k-Series and O2k-Series E or higher. |
| Dissection Set | | The Oroboros Dissection Set: for tissue preparation, set of 4 pairs of stainless steel, antimagnetic forceps and a pair of scissors. |
| Electrolyte\Reference-Electrode | | Electrolyte\Reference-Electrode for Reference-Electrode\2.4 mm |
| Filter Set AmR | | Filter Set AmR: Set of filters for the determination of H2O2 production with 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 | | Filter set MgG / CaG: Set of filters for the determination of concentraions of Mg2+ or Ca2+ with the fluorophores 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 | | Filter set Saf: Set of filters for the (qualitative) determination of mitochondrial membrane potential with 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). |
| Filter-Cap | | Filter-Cap: O2k-Fluo LED2-Module (O2k-Series D to G) sensors (Fluorescence-Sensor Green and Fluorescence-Sensor Blue) and Oroboros 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. |
| Fluo LED2-Module | | The Fluo LED2-Module is a component of the Oroboros FluoRespirometer (O2k-Series D to G). It is an amperometric add-on module to O2k-Series D to G, adding a new dimension to high-resolution respirometry. Optical sensors are inserted through the front window of the Oroboros 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 Oroboros user innovation. |
| Fluo Smart-Module | | The Oroboros Fluo Smart-Module is an amperometric add-on module to the Oroboros, adding a new dimension to high-resolution respirometry. Optical sensors are inserted through the front window of the Oroboros 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 Oroboros user innovation.
- » MiPNet28.09 Fluo Smart-Module manual
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| Fluorescence-Control Unit | | Fluorescence-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 Fluo LED2-Module. |
| Fluorescence-Sensor Blue | | Fluorescence-Sensor Blue: excitation LED 465 nm (dominant wavelength), 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. |
| Fluorescence-Sensor Green | | Fluorescence-Sensor Green: excitation LED 525 nm (dominant wavelength), photodiode, Filter-Cap equipped with Filter Set AmR for Amplex UltraRed measurements when delivered. |
| Forceps for membrane application | | Forceps for membrane application: for OroboPOS and ISE membrane application; do not use for tissue preparation. |
| Forceps\stainless Steel\angular Tip\fine | | Forceps\stainless Steel\angular Tip\fine: for tissue preparation, stainless steel. Two pairs are used particularly for muscle fiber separation. |
| Forceps\stainless Steel\rounded Tip\sharp | | Forceps\stainless Steel\rounded Tip\sharp: for 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 | | Forceps\stainless Steel\straight Tip\sharp: for tissue preparation, stainless steel, antimagnetic. One pair is recommended for insertion of the sample into the Oroboros Chamber and for handling in combination with Forceps\stainless Steel\rounded Tip\sharp. |
| High-resolution respirometry | HRR | 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 with the Oroboros Bioenergetics Analyzer combined with MultiSensor modules.
HRR provides the basis for precision OXPHOS analysis. Mitochondrial function and dysfunction have gained increasing interest, reflecting growing awareness of the fact that mitochondria play a pivotal role in human health and disease. HRR combines instrumental accuracy and reliability with the versatility of substrate-inhibitor-uncoupler titrations: SUIT protocols allow the interrogation of numerous mitochondrial pathway and coupling control states in a single respirometric assay. Precision OXPHOS analysis displays respiratory pathway and coupling control patterns and reveals a diversity of bioenergetic profiles in mitochondrial comparative physiology and diagnostic applications.
Technologically, HRR is based on the Oroboros Bioenergetics Analyzer (Oroboros), 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). The Oroboros provides sole source instruments, with no other available platform meeting their specifications for high-resolution respirometry. 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 to MultiSensor analysis. Smart Fluo-Sensors are integrated into the Oroboros to measure simultaneously fluorometric signals using specific fluorophores. Potentiometric modules are available with ion-selective electrodes (pH, TPP+). The PB-Module extends HRR to PhotoBiology with accurate control of a wide range of light intensities and measurement of photosynthesis.
The Oroboros O2k and the Oroboros NextGen-O2k support all these Oroboros Modules. The NextGen-O2k all-in-one, however, is unique in supporting Q-Redox and NADH-Redox Modules. |
| Hydrogenion flux | JH+ | Volume-specific hydrogenion flux or H+ flux is measured in a closed system as the time derivative of H+ concentration, expressed in units [pmol·s-1·mL-1]. H+ 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 H+ 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 hydrogen ion activity. Therefore, ECAR is of interest in relation to acidification issues in the incubation buffer or culture medium. The physiologically relevant metabolic H+ flux, however, must not be confused with ECAR. |
| ISS-Waste Bottle | | ISS-Waste Bottle, 2-liter, component of the ISS-Integrated Suction System. |
| MiR05-Kit | | Mitochondrial Respiration Medium - MiR05-Kit, 1 vial; for a final volume of 250 mL |
| Microsyringe\10 mm3 51/0.13 mm | | Hamilton Microsyringe\10 mm3 51/0.13 mm for manual titrations, 10 mm3 volume; fixed injection needle with rounded tip: 51 mm length, 0.13 mm inner diameter. |
| Microsyringe\100 mm3 51/0.41 mm | | Hamilton Microsyringe\100 mm3 51/0.41 mm for manual titrations, 100 mm3 volume; fixed needle with rounded tip: 51 mm length, 0.41 mm inner diameter. It is recommended for injections of suspensions of isolated mitochondria. |
| Microsyringe\200 mm3\TIP2k | | Microsyringe\200 mm3\TIP2k: Microinjection syringe for Titration-Injection microPump, 200 mm3 (µl), fixed injection needle with rounded tip, with spacers. |
| Microsyringe\25 mm3 51/0.15 mm | | Hamilton Microsyringe\25 mm3 51/0.15 mm for manual titrations, 25 mm3 volume; fixed needle with rounded tip: 51 mm length, 0.15 mm inner diameter. |
| Microsyringe\50 mm3 51/0.15 mm | | Hamilton Microsyringe\50 mm3 51/0.15 mm for manual titrations, 50 mm3 volume; fixed needle with rounded tip: 51 mm length, 0.15 mm inner diameter. |
| Microsyringe\500 mm3\TIP2k | | Microsyringe\500 mm3\TIP2k: Microinjection syringe for Titration-Injection microPump, 500 mm3 (µl), fixed injection needle with rounded tip, with spacers. |
| O2k-Fluorometer Series G | | O2k-FluoRespirometer Series G - Former Series (up to 2017-July) - the experimental system complete for high-resolution respirometry (HRR) combined with fluorometry. The O2k-Fluorometer G includes the Oroboros Main Unit, Fluo LED2-Module and TIP2k, and supports all other add-on O2k-Modules.
The Oroboros is a sole source apparatus with no other instruments meeting its specifications. |
| O2k-Window Frame | |
For O2k-Series D - G: O2k-Window Frame: blue POM, with thread for fixation on the Oroboros 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 Tool | |
O2k-Window Tool for removing the blue O2k-Window Frame from the Oroboros Main Unit, for rare cleaning purposes and for front fixation of the Fluorescence-Control Unit. |
| Oroboros | |
The Oroboros is the Bioenergetics Platform for high-resolution respirometry (HRR) for mitochondria and cell research, unique for precision OXPHOS analysis.
The Oroboros is available as two models:
- Oroboros NextGen-O2k
- Oroboros O2k
Both Oroboros models provide the basis for HRR applications combined with measurement of ROS production, mitochondrial membrane potential, ATP production, Ca2+, pH, and PhotoBiology.
The modular Oroboros concept is supported by DatLab, with high flexibility for add-on Modules. All Oroboros Modules are supported by the Oroboros NextGen-O2k, which is the only model supporting the Redox Modules (Q-Module and NADH-Module), and the NO-Module.
The Oroboros allows bioenergetic analysis at controlled oxygen levels. 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 globally tested and trusted high-resolution Oroboros prioritizes both quality and scientific research output in the field of mitochondrial physiology and pathology, extended to PhotoBiology. The Oroboros is a robust sole-source instrument with a long life time and low running costs. Expert service is provided by the Oroboros Open Support. |
| Oroboros Chamber | |
Oroboros Chamber: Duran® glass polished, with experimental volumes (V) of 2.0 mL (classic volume cV, 16 mm inner diameter) or 0.5 mL (small volume sV, sV-Module, 12 mm inner diameter). The optical properties of Duran® allow application of fluorometric sensors (Duran® optical properties). |
| Oroboros FluoRespirometer | | The Oroboros FluoRespirometer - the Bioenergetics Platform for high-resolution respirometry (HRR) and fluorometry, allows 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 Oroboros supports the add-on modules: ISE TPP-Module, ISE pH-Module, enabling measurement of mt-membrane potential with ion sensitive electrodes (ISE for TPP+ or TPMP+) or pH. |
| Oroboros Main Unit | | The Oroboros Main Unit is a component of the Oroboros O2k or the NextGen-O2k (» more information). The Oroboros Main Unit consists of functionally defined, integral elements, the (Oroboros Main Unit, Oroboros Peltier Temperature Control, two O2k-Electromagnetic Stirrer Twin-Control units, two O2k-Amperometric OroboPOS Twin-Channels, Oroboros Barometric Pressure Transducer), which cannot be obtained separately. |
| Oroboros Titration Set | | The Oroboros 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. |
| Oroboros specifications | | Oroboros versus multiwell respirometer: The Oroboros Bioenergetics Platform for high-resolution respirometry, the Oroboros O2k and Oroboros NextGen-O2k meet the powerful quality criteria securing high output and pioneering state-of-the-art precision OXPHOS analysis of substrate control and coupling control of mitochondrial function. The limit of detection of O2 flux is as low as 0.5 pmol·s-1·mL-1. Signal noise at zero oxygen concentration is <0.05 μM O2. Oxygen backdiffusion at zero oxygen is <3 pmol·s-1·mL-1, and oxygen consumption at air saturation and standard barometric pressure (100 kPa) was 2.7 ± 0.9 SD in 114 test runs at 37 °C. These highly standardized instrumental background fluxes are a linear function of oxygen concentration, which is used for routine background correction of oxygen flux. Typical exponential time constants of the oxygen sensors are <4 s, used for dynamic corrections in kinetic studies.
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). High throughput stands for multiwell systems - expensive, with limited scope and high running costs. In respirometry, high throughput is not equivalent to high output. High output is the concept of the two-chamber Oroboros O2k and NextGen-O2k. |
| Power O2k Numbers | | Power O2k Numbers: Single number to label the O2k in a Power O2k-Lab. |
| 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 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 Oroboros Titration Set. Furthermore, the OroboPOS-Mounting Tool of the OroboPOS Service Tools can be used from the available O2k and is not included. |
| Smart Fluo-Sensor Blue | | Smart Fluo-Sensor Blue: Excitation LED 465 nm (dominant wavelenght) with short pass filter, emission (red) with long pass filter, individual sensors are calibrated with sensor-specific memory and direct input into DatLab 7 to obtain reproducible light intensities with different sensors, including photodiode, 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 | | Smart Fluo-Sensor Green: Excitation LED 525 nm (dominant wavelength) with short pass filter, emission (red) with long pass filter, individual sensors are calibrated with sensor-specific memory and direct input into DatLab 7 to obtain reproducible light intensities with different sensors, including photodiode, Filter-Cap equipped with Filter Set AmR for Amplex UltraRed and TMRM measurements when delivered. |
| Troubleshooting: fluorescence module | | |
