Difference between revisions of "High-resolution respirometry"

Revision as of 09:42, 29 March 2024

high-resolution terminology - matching measurements at high-resolution

High-resolution respirometry

Description

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 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 applicable protocols, allowing practically unlimited addition and combination of substrates, inhibitors, and uncouplers in the Oroboros O2k. Substrate-uncoupler-inhibitor titration (SUIT) protocols allow the interrogation of numerous mitochondrial pathway and coupling states in a single respirometric assay. Mitochondrial respiratory pathways may be analyzed in detail to evaluate even minor alterations in respiratory coupling and pathway control patterns.

The O2k is a sole source technology, with no other available instrument meeting its specifications for high-resolution respirometry. 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-Fluo Smart-Module. 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 PB-Module extends HRR to PhotoBiology with accurate control of the light intensity and measurement of photosynthesis. The O2k-J and the NextGen-O2k support all these O2k-Modules. The NextGen-O2k all-in-one, however, is unique in supporting Q-Redox and NADH-Redox Modules.

Abbreviation: HRR

Reference: Oroboros O2k

Power O2k: Several O2k units are operated simultaneously (P1, P2, ..) to combine high-resolution respirometry with high-output.

Template:Keywords: O2k

MitoPedia: O2k and high-resolution respirometry

» MitoPedia: O2k hardware

» MitoPedia: DatLab

» MitoPedia: SUIT

Links

» MitoPedia: Respiratory states

-

» Respirometry

» MitoPedia: Respirometry

» Wikipedia: Respirometry

» O2k Applications

» Keywords

Instrument

» DatLab; » MitoPedia: DatLab

» Instrumental background oxygen flux »» MiPNet14.06 Instrumental O2 background

» Oroboros O2k-Core (O2k-Series D - G)

» O2k-FluoRespirometer

» O2k-MultiSensor

» Oroboros O2k; » MitoPedia: O2k hardware

» Power O2k-Respirometer

» Static sensor calibration

» TIP2k

Concept

» Concentration

» Coupling-control state

» High-resolution respirometry

» Mitochondrial preparations

» Oxidative phosphorylation

» Oxygen, dioxygen, O₂

» Oxygen flux

» Electron-transfer-pathway state

» Respiration

» Respirometry; » MitoPedia: Respirometry

» Substrate-uncoupler-inhibitor titration (SUIT) protocols; » MitoPedia: SUIT

» References

O2k-Publications

Keywords: Oxygen signal

Bioblast links: Oxygen signal - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>

Specific

O2k-Procedures

» High signal at zero oxygen

» POS calibration - static

» POS calibration - dynamic

MiPNet O2k-Procedures

» POS-calibration

» O2k-Manual: POS-service

» O2k-cleaning

» O2k-Series G and DatLab 6: Calibration

General

» Oxygen, dioxygen, O₂

» Oxygen calibration - DatLab

» Oxygen solubility

» Oxygen solubility factor

» Oxygen pressure

» Concentration

» Activity

» Pressure - Pascal

» Barometric pressure

» High-resolution respirometry

» OroboPOS

» Polarographic oxygen sensor

» MitoFit Quality Control System

» Sensitivity

Other keyword lists

» Keywords: Concentration and pressure

Selected references

MitoPedia methods: Respirometry, Fluorometry, Spectrophotometry

MitoPedia O2k and high-resolution respirometry: O2k hardware, O2k-Respirometry, O2k-FluoRespirometry

@@ Line 1: / Line 1: @@
 {{MitoPedia
 |abbr=HRR
-|description=[[Image:O2k-Fluorometer.JPG|180px|left|link=O2k-FluoRespirometer]]
+|description=[[Image:O2k-Fluorometer.jpg|200px|left|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. The most advanced way to analyze mitochondrial function is by means of '''High-Resolution Respirometry''' (HRR) and [[High-Resolution FluoRespirometry]] (HRFR) using the [[Oroboros O2k]]. 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-Module|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. The most important extension of the [[Startup O2k-Respirometer |O2k-Respirometer]] is the [[O2k-FluoRespirometer]].
-'''High-Resolution Respirometry''' (HRR) is defined on the basis of instrumental features of the O2k. The limit of detection of oxygen flux is as low as 1 pmol O<sub>2</sub>.s<sup>-1</sup>.ml<sup>-1</sup>. Oxygen backdiffusion at zero oxygen is <4 pmol.s<sup>-1</sup>.ml<sup>-1</sup>. Oxygen consumption by the polarographic oxygen sensor at air saturation and standard barometric pressure (100 kPa) is 2.7 pmol.s<sup>-1</sup>.ml<sup>-1</sup> (± 0.9 SD; 114 test runs at 37 °C). These highly standardized instrumental background fluxes are a linear function of oxygen concentration, which is used for automatic background correction of oxygen flux (DatLab). Typical exponential time constants of the oxygen sensors are <4 s, used for dynamic corrections in kinetic studies. Signal noise at zero oxygen concentration is <0.05 µM O<sub>2</sub>.
+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 applicable protocols, allowing practically unlimited addition and combination of substrates, inhibitors, and uncouplers in the [[Oroboros O2k]]. Substrate-uncoupler-inhibitor titration (SUIT) protocols allow the interrogation of numerous mitochondrial pathway and coupling states in a single respirometric assay. Mitochondrial respiratory pathways may be analyzed in detail to evaluate even minor alterations in respiratory coupling and pathway control patterns.
-|info=[[Gnaiger 1995 J Bioenerg Biomembr]]
-}}
+The O2k is a sole source technology, with no other available instrument meeting its specifications for high-resolution respirometry. 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-Module|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-Fluo Smart-Module]]. [[Smart Fluo-Sensor]]s are integrated into the O2k to measure simultaneously fluorometric signals using specific fluorophores. Potentiometric modules are available with ion-selective electrodes (pH, TPP<sup>+)</sup>. The [[PB-Module]] extends HRR to PhotoBiology with accurate control of the light intensity and measurement of photosynthesis. The O2k-J and the NextGen-O2k support all these O2k-Modules. The [[NextGen-O2k]] all-in-one, however, is unique in supporting Q-Redox and NADH-Redox Modules.
-{{MitoPedia methods
+|info=[[Oroboros O2k]]
-|mitopedia method=Respirometry, Fluorometry, Spectrophotometry
-}}
-{{MitoPedia O2k and high-resolution respirometry
-|mitopedia O2k and high-resolution respirometry=DatLab
 }}
+::::* <big>'''Power O2k''':</big> Several O2k units are operated simultaneously ('''P'''1, '''P'''2, ..) to combine high-resolution respirometry with [[Power O2k-Respirometer |high-output]].
+{{Template:Keywords: O2k}}
+== MitoPedia: O2k and high-resolution respirometry ==
+::::» [[MitoPedia: O2k hardware]]
+::::» [[MitoPedia: DatLab]]
+::::» [[MitoPedia: SUIT]]
+== Links ==
+::::'''»''' [[MitoPedia: Respiratory states]] [[Image:P.jpg|link=OXPHOS capacity|OXPHOS]] [[Image:R.jpg|link=ROUTINE respiration|ROUTINE]] [[Image:E.jpg|link=ET capacity|ET capacity]] [[Image:L.jpg|link=LEAK respiration|LEAK]]  - [[Image:ROX.jpg|link=Residual oxygen consumption|ROX]]
+::::» [[Respirometry]]
+::::» [[MitoPedia: Respirometry]]
+::::» [http://en.wikipedia.org/wiki/Respirometry Wikipedia: Respirometry]
+::::» [[O2k Applications]]
 <br />
 [[File:Expand.png|right|45px |Click to expand or collaps]]
@@ Line 21: / Line 34: @@
 ::::» [[DatLab]]; » [[MitoPedia: DatLab]]
 ::::» Instrumental background oxygen flux »» [[MiPNet14.06 Instrumental O2 background]]
-::::» [[O2k-Core]]
+::::» [[Oroboros O2k-Core (O2k-Series D - G)]]
 ::::» [[O2k-FluoRespirometer]]
 ::::» [[O2k-MultiSensor]]
 ::::» [[Oroboros O2k]]; » [[MitoPedia: O2k hardware]]
-::::» [[Power-O2k]]
+::::» [[Power O2k-Respirometer]]
 ::::» [[MiPNet06.03 POS-calibration-SOP |Static sensor calibration]]
 ::::» [[TIP2k-Module|TIP2k]]
@@ Line 31: / Line 44: @@
 ::: '''Concept'''
 ::::» [[Concentration]]
-::::» [[Coupling control state]]
+::::» [[Coupling-control state]]
-::::» [[High-Resolution FluoRespirometry]]
+::::» [[High-resolution respirometry]]
 ::::» [[Mitochondrial preparations]]
 ::::» [[Oxidative phosphorylation]]
 ::::» [[Oxygen |Oxygen, dioxygen, O<sub>2</sub>]]
 ::::» [[Oxygen flux]]
-::::» [[Pathway control state]]
+::::» [[Electron-transfer-pathway state]]
 ::::» [[Respiration]]
 ::::» [[Respirometry]]; » [[MitoPedia: Respirometry]]
@@ Line 44: / Line 57: @@
 </div>
 <br />
-[[Image:O2.jpg|left|60px|link=Oxygen|Dioxygen]]
-== MitoPedia: O2k ==
-::::» [[MitoPedia: O2k hardware]]
-::::» [[MitoPedia: DatLab]]
-::::» [[MitoPedia: SUIT]]
-== Links ==
-::::'''»''' [[MitoPedia: Respiratory states]] [[Image:P.jpg|link=OXPHOS capacity|OXPHOS]] [[Image:R.jpg|link=ROUTINE respiration|ROUTINE]] [[Image:E.jpg|link=ETS capacity|ETS]] [[Image:L.jpg|link=LEAK respiration|LEAK]]  - [[Image:ROX.jpg|link=Residual oxygen consumption|ROX]]
-::::» [[Respirometry]]
-::::» [[MitoPedia: Respirometry]]
-::::» [http://en.wikipedia.org/wiki/Respirometry Wikipedia: Respirometry]
-::::» [[O2k Applications]]
 [[Image:O2k-Publications.jpg|left|116px|link=O2k-Publications: Topics|O2k-Publications in the MiPMap]]
@@ Line 71: / Line 67: @@
 ::::* [[O2k-Publications |O2k-Publications: Chronological]]
 ::::* [[O2k-Publications: Topics]]
+== Keywords: Oxygen signal ==
+{{Template:Keywords: Oxygen signal}}
 ::: '''Selected references'''
@@ Line 76: / Line 77: @@
 ::::* [[Gnaiger 2001 Respir Physiol]]
 ::::* [[Gnaiger 2008 POS]]
-::::* [[Gnaiger 2014 MitoPathways]]
+::::* [[Gnaiger 2020 BEC MitoPathways]]
 </div>
 </div>
 <br />
+{{MitoPedia methods
+|mitopedia method=Respirometry, Fluorometry, Spectrophotometry
+}}
+{{MitoPedia O2k and high-resolution respirometry
+|mitopedia O2k and high-resolution respirometry=O2k hardware, O2k-Respirometry, O2k-FluoRespirometry
+}}