MitoPedia: O2k-Respirometry

From Bioblast
Jump to: navigation, search

Header ecosystem 2020.jpg

OROBOROS support
        MitoPedia: O2k-Open Support         MitoPedia: O2k hardware         MitoPedia: DatLab         MitoPedia: SUIT         O2k-Procedures         O2k-Manual         O2k-Videosupport        

MitoPedia: O2k-Respirometry

Template NextGen-O2k.jpg
O2k-Open Support

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

MitoPedia: O2k-Respirometry is a component of O2k-Open Support.
It lists basic HRR-related topics.
See also
» MitoPedia: O2k-FluoRespirometry

Cell count and normalization in HRRNceThe 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 Preprint Arch 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).
Comparison of respirometric methodsThe comparison of respirometric methods provides the basis to evaluate different instrumental platforms and different mitochondrial preparations, as a guide to select the best approach and to critically evaluate published results.
Cover-Slip black.JPG
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. Two units of this item are standard components of the O2k-Assembly Kit.
High and unstable zero current of oxygen sensorR0High and unstable zero current of oxygen sensor
High-resolution respirometryHRR
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.
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.
Mitochondrial Respiration Medium - MiR05-Kit, 1 vial; for a final volume of 250 mL
Microbalance 120 g
Microbalance 120 g.jpg
Microbalance 41 g to 120 g; 0.01 mg / 0.1 mg display; particularly for wet weight determination of permeabilized fibres.
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).
Motic Microscope
Motic Microscope.jpg
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\Viton\12.5x1 mm
O-ringViton12.5x1 mm.jpg
O-ring\Viton\12.5x1 mm, for PVDF or PEEK O2k-Stoppers (2-mL O2k-Chamber), box of 8 as spares.

Two spare boxes of this product are standard components of the O2k-Assembly Kit (O2k-FluoRespirometer) as well as the O2k-TPP+ ISE-Module, O2k-pH ISE-Module and the O2k-NO Amp-Module.

Two O-rings (O-ring\Viton\12.5x1 mm) are mounted on each PVDF or PEEK O2k-Stopper. The spare O-rings are placed in the O2k-Service Box.
O2k-Dissection Set
O2k-Dissection Set: for tissue preparation, set of 4 pairs of stainless steel, antimagnetic forceps and a pair of scissors.
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-Titration Set
O2k-Titration Set.JPG
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-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).
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.
POS connector and cable connection.jpg
OroboPOS-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.
Pos holder.JPG
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.
OroboPOS-Service Kit
ServiceBox G 2.jpg
OroboPOS-Service_Kit, with O2k-Service Box including all oxygen sensor service accessories for membrane mounting and service of the OroboPOS.
PBI-Shredder O2k-Set
PBI-Shredder HRR-Set.JPG
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.
Packing\Peli Case
Packing Peli Case.jpg
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)
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
Power O2k notext.png
The Power O2k-Respirometer is an economical option for using additional O2k-Units to increase output in high-resolution respirometry.
Screwdriver allen wrench
Screwdriver allen wrench, a standard component of the O2k-FluoRespirometer and O2k-sV-Module.
Startup O2k-Respirometer
Startup O2k-Respirometer notitle.png
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\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.

Two units of this item are standard components of the O2k-Assembly Kit (O2k-FluoRespirometer).

For dispatch of the O2k-FluoRespirometer, the Stirrer-Bars are shipped in the O2k-Service Box.
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. Two units of this item are standard components of the O2k-Assembly Kit.
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).
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.