Difference between revisions of "High-resolution respirometry"

Latest revision as of 09:09, 2 April 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 using the Oroboros O2k-technology. 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-technology provides sole source instruments, with no other available instrument meeting its specifications for high-resolution respirometry. Technologically, HRR is based on the Oroboros O2k-technology, 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 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-technology

»Bioblast links: O2k-technology and HRR - >>>>>>> - Click on [Expand] or [Collapse] - >>>>>>>

O2k-technology

» Oroboros O2k-technology

» High-resolution respirometry

» NextGen-O2k - Category:NextGen-O2k - NextGen-O2k Technical developments

» Oroboros O2k

» Power O2k-Respirometer

» MitoPedia: O2k hardware

» DatLab; » MitoPedia: DatLab

» O2k-MultiSensor, O2k-FluoRespirometer

Oroboros Marketplace

» O2k

Concept

» Concentration

» Coupling-control state

» 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

MitoPedia: O2k and high-resolution respirometry

» MitoPedia: O2k hardware

» MitoPedia: DatLab

» MitoPedia: SUIT

» MitoPedia: Respiratory states

-

» Respirometry

» MitoPedia: Respirometry

» Wikipedia: Respirometry

» O2k Applications

O2k-Publications

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-Fluorometer]]
+|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 ten years, reflecting growing awareness of the fact that mitochondria play a pivotal role in human health and disease. The most suitable way to analyze mitochondrial function is by means of '''Oxygraph-2k''' (O2k) '''high-resolution respirometry''' applying [[SUIT| substrate-uncoupler-inhibitor titration (SUIT) protocols]] that allow the diagnosis of numerous mitochondrial pathway defects in a single respirometry assay. Technologically, HRR is based on the [[Oxygraph-2k|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 ([[MiPNet06.03 POS-calibration-SOP |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 to the [[O2k-MultiSensor]] systems is the [[O2k-Fluorometer]].
-|info=[[Gnaiger 1995 J Bioenerg Biomembr]], [[Gnaiger 2008 POS]], [[Gnaiger 2001 Respir Physiol]], [[Gnaiger 2014 MitoPathways]]
-}}
-{{MitoPedia concepts}}
-{{MitoPedia methods
-|mitopedia method=Respirometry, Fluorometry, Spectrophotometry
-}}
-{{MitoPedia O2k and high-resolution respirometry
-|mitopedia O2k and high-resolution respirometry=DatLab
-}}
-{{MitoPedia topics}}
-[[Image:O2.jpg|left|60px|link=Oxygen|Dioxygen]]
-== What is an Oxygraph? ==
-::::* <big>'''O2'''k:</big> The [[Oxygraph-2k |O2k]] measures oxygen ([[Oxygen |dioxygen, '''O<sub>2</sub>''']]) as a gas dissolved in aqueous solution.
-::::* With the O2k, experimental data are displayed not only of oxygen concentration over time; but real-time transformation of these data allows the experimenter to follow '''respiration''' (oxygen consumption, oxygen flux) of biological and biochemical samples while the experiment is still running.
-::::* An Oxygraph is not used to measure oxygen concentration in a tissue, but for high-resolution analysis of respiratory activity of biological or biochemical samples transferred into the experimental chamber.
-[[Image:O2k-Protocols.jpg|right|230px|link=Gnaiger_2014_MitoPathways|MitoPathways]]
-[[File:Pesta 2012 Methods Mol Biol Fig7A.jpg|left|250px|link=Pesta_2012_Methods_Mol_Biol|Pesta and Gnaiger 2012]]
-:::: Specific [[substrate-uncoupler-inhibitor titration]] (SUIT) protocols are applied in studies of cell respiration or mitochondrial activity. In experiments with intact cells, different [[Coupling control state|coupling states]] can be studied to evaluate ROUTINE respiratory activity, respiration coupled to ATP production, respiratory excess capacity, residual oxygen consumption, and, by extension of conventional protocols, intactness of cell membranes. In [[Mitochondrial preparations|mitochondrial preparations]] such as isolated mitochondria, permeabilized cells and tissue samples, or homogenized tissue, quantitative information is obtained on selected [[Gnaiger 2014 MitoPathways |mitochondrial pathways]] is obtained, particularly of the process of '''[[oxidative phosphorylation]] (OXPHOS)''' in mitochondria and bacteria. Comprehensive OXPHOS analysis leads us beyond the highly informative process of oxygen consumption, when high-resolution respirometry is combined with [[O2k-MultiSensor]] modules to simultaneously measure the production of reactive oxygen species or mitochondrial membrane potential, nitric oxide production, acid-base balance or calcium with O2k-Fluorometry or ion selective electrodes. In the O2k, these bioenergetic parameters are always evaluated in combination with the measurement of oxygen concentration and respiration.
-[[File:O2k-universe.jpg|right|240px|]]
-::::* <big>O'''2k''':</big> The O2k is the '''2'''<sup>nd</sup> generation of our high-resolution respirometers. The first model was available as of [[OROBOROS 1992|1992]]. Development of the new O'''2k''' started in '''2k''' (2000) and the first O'''2'''k specimen was delivered in [[OROBOROS 2002|'''2'''00'''2''']].
-::::* <big>O2'''k''':</big> The O2k has two experimental chambers ('''k''': Kammern, German for chambers). [[O2k-Core |Why 2 ml?]]
-::::* <big>'''Power-O2k''':</big> Several O2k units are operated simultaneously ('''P'''1, '''P'''2, ..) to combine high-resolution respirometry with [[Power-O2k |high-output]].
+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 using the [[Oroboros O2k-technology]]. 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-technology provides sole source instruments, with no other available instrument meeting its specifications for high-resolution respirometry. Technologically, HRR is based on the Oroboros O2k-technology, 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 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.
-::::* '''<big>[[OROBOROS INSTRUMENTS|O2k by OROBOROS - tested and trusted]]</big>'''
+|info=[[Oroboros O2k-technology]]
+}}
+{{Template:Keywords: O2k-technology}}
 == MitoPedia: O2k and high-resolution respirometry ==
@@ Line 55: / Line 15: @@
 ::::» [[MitoPedia: SUIT]]
+::::'''»''' [[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]]
-== 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]]
+::: '''O2k-Publications'''
+::::* [[O2k-Publications |O2k-Publications: Chronological]]
+::::* [[O2k-Publications: Topics]]
+{{Template:Keywords: Oxygen signal}}
+::: '''Selected references'''
+::::* [[Gnaiger 1995 J Bioenerg Biomembr]]
+::::* [[Gnaiger 2001 Respir Physiol]]
+::::* [[Gnaiger 2008 POS]]
+::::* [[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
+}}