Search results

...[[SUIT-003]]''' Coupling-control protocol and respirometric cell viability test (CCV protocol) without oligomycin. ::: '''[[Categories of SUIT protocols |SUIT-category]]:''' cePMGlc,S

|info='''C''' - '''[[SUIT-003]]''' |SUIT number=D013_ce1;ce1P;ce2Omy;ce3U;ce3Glc;ce3M;ce4Rot;ce5S;1Dig;1U;1c;2Ama;3A

|info='''B''': '''[[SUIT-018]]''' |SUIT number=D054_mt;1GMS;2D;2c;3Ama

|abbr=Digitonin test |description=[[File:ce1;ce2Rot;ce3S;ce4D;1Dig;1c.png|400px|Respirometric test of optimum digitonin concentration ]]

::::# [[MitoFit protocols]] ::::## [[Harmonization protocols]] (Begley, Ioannidis 2015)

|info='''A''' Coupling-control protocol and Crabtree effect test -'''[[SUIT-003]]''' complex cell [[coupling-control protocol]] |SUIT number=D038_ce1;(ce2Omy);ce3U;ce3Glc;ce3'U;ce4Ama

|info='''B''' - [[SUIT-003]] |SUIT number=D012_ce1;ce1P;ce2Omy;ce3U;ce4Rot;ce5Ama}}

|description=[[File:ce1;ce2Rot;ce3S;ce4D;1Dig;1c.png|400px|Respirometric test of optimum digitonin concentration ]] |info='''A''' -'''[[SUIT-010]]''' - Optimum [[digitonin]] concentration for [[permeabilized cells |p

|description=[[File:SUIT-003 O2 ce D067 diagram.png|350px]] ...ontrol protocol]] and [[Crabtree effect]] with glycolysis inhibition- '''[[SUIT-003]]'''

|info='''A''' '''[[SUIT-003]]''' - [[Coupling-control protocol]] and [[Crabtree effect]] |SUIT number=D037_ce1;ce1Glc;(ce2Omy);ce3U;ce4Ama

...-003]]''' - [[Coupling-control protocol]] and respirometric cell viability test; ce: living cells, pce: permeabilized cells |SUIT number=D020_ce1;ce1P;ce2Omy;ce3U;ce4Rot;ce5S;1Dig;1c;2Ama;3AsTm;4Azd

...version of the Coupling-control protocol and respirometric cell viability test '''(CCV protocol)''' |SUIT number=D018_ce1;ce1P;ce2Omy;ce3U;ce3Glc;ce4Rot;ce5S;1Dig;1c;2Ama;3AsTm;4Azd

...s (isolated mitochondria, tissue homogenate and permeabilized cells)- '''[[SUIT-032]]''' |SUIT number=D109_mt;1PGM;2D;3Myx

...the Q-pool in S(Rot)-pathway control state in isolated mitochondria- '''[[SUIT-006]]''' |SUIT number=D071_mt;1Q2;1Rot;1S;2D;(3Omy);4U;5Anox;6Ama.png

...s (isolated mitochondria, tissue homogenate and permeabilized cells)- '''[[SUIT-006]]''' |SUIT number=D108_mt;1PGM;2D;2c;(3Omy);4U;5Myx

...trol in intact cells without oligomycin; ce: non-permeabilized cells -'''[[SUIT-003]]''' |SUIT number=D039_ce1;(ce2Omy);ce3U;ce4Rot;ce5Ama

...s (isolated mitochondria, tissue homogenate and permeabilized cells)- '''[[SUIT-034]]''' |SUIT number=D082_mt;1D.1;2PGM;3D2.5;4U;5Myx

|info='''B:''' Experimental control for [[SUIT-003 AmR ce D058]] |SUIT number=D059_ce1;ce1H2O;ce1MiR05;ce1DMSO;ce2Omy;ce3U;ce4Rot;ce5Ama

|abbr=Optimum [acylcarnitine] test |info='''A''' - '''[[SUIT-041]]''' - '''[[Fatty_acid_oxidation_pathway_control_state|F-pathway]] an

...s (isolated mitochondria, tissue homogenate and permeabilized cells)- '''[[SUIT-033]]''' |SUIT number=D110_mt;1D.1;2PGM;3D2.5;4Myx

|info='''A''' -'''[[SUIT-018]]''' - SUIT protocol for simultaneous determination of O₂ and H₂O |SUIT number=D068_ce1;1Dig;1GMS;2D;3Ama

...control in high-resolution respirometry, aimed at standardization of SUIT protocols and fostering proficiency to generate experimental data of assured high qua ...using standardized mitochondrial reference samples, DatLab analysis, SUIT protocols and experimental design, normalization of flux and data reporting, O2k-Fluo

...of the Q-pool in S(Rot)-pathway control state in permeabilized cells- '''[[SUIT-006]]''' |SUIT number=D073_ce1;1Dig;1Q2;1Rot;1S;2D;(3Omy);4U;5Anox;6Ama.png

...compounds [[MitoKit-CII/Succinate-nv]] and [[MitoKit-CII/Malonate-nv]] - [[SUIT-003 O2 ce D060]] |SUIT number=D061_ce1;(ce2Omy);ce3U;ce4Rot;ce5DMSO;ce6DMSO;ce7Ama

|abbr=CCP-ce S permeability test |info='''B''': CCP(P) and cell membrane permeability test with succinate - [[SUIT-003]]

|info='''A''' -'''[[SUIT-018]]''' |SUIT number=D041_mt;1GMS;2D;3Ama

...mR assay components on the respiration ; ce: non-permeabilized cells -'''[[SUIT-003]]''' |SUIT number=D058_ce1;ce1SOD;ce1HRP;ce1AmR;ce2Omy;ce3U;ce4Rot;ce5Ama

...s (isolated mitochondria, tissue homogenate and permeabilized cells)- '''[[SUIT-032]]''' |SUIT number=D078_File:1mt;1PGM;2D;3Anox;4Myx;5Reox

...mpounds [[MitoKit-CII/Succinate-nv]] and [[MitoKit-CII/Malonate-nv]] -'''[[SUIT-003]]''' - Coupling-control protocol (CCP) with plasma membrane-permeable s |SUIT number=D060_ce1;(ce2Omy);ce3U;ce4Rot;ce5Snv;ce6Mnanv;ce7Ama

...s (isolated mitochondria, tissue homogenate and permeabilized cells)- '''[[SUIT-033]]''' |SUIT number=D081_mt;1D.1;2PGM;3D2.5;4Anox;5Myx;6Reox

...s (isolated mitochondria, tissue homogenate and permeabilized cells)- '''[[SUIT-034]]''' |SUIT number=D082_mt;1D.1;2PGM;3D2.5;4U;5Anox;6Myx;7Reox

::: '''[[SUIT protocol pattern]]:''' 1ce;2ceOmy;3ceU;ce*, or ce1;ce2Omy;ce3U;ce4Rot;ce5S; ...dular approach and can be easily used in combination with the permeability test and/or the assessment of the complex IV capacity.

...s (isolated mitochondria, tissue homogenate and permeabilized cells)- '''[[SUIT-006]]''' |SUIT number=D048_1PM;2D;3Omy;4U;5Ama

::: '''[[SUIT protocol pattern]]:''' 1OctM;2D;3G;4P;5S;6U:7Rot- ...]] and of [[FNS]] and [[S]] in [[ET capacity| ET state]] can be evaluated. SUIT-015 can be extended with the CIV assay module.

...on=[[File:Pfi;1PM;2D;2c;3U;4G;5S;6Oct;7Rot;8Gp;9Ama;10AsTm;11Azd.png|400px|SUIT-RP1]] |info=[[SUIT-001]] - '''[[SUIT reference protocol]] Reference protocol RP1 for permeabilized fibers'''

...s (isolated mitochondria, tissue homogenate and permeabilized cells)- '''[[SUIT-006]]''' |SUIT number=D084_mt;1PGM;2D;3(Omy);4U;5Anox;6Myx;7Reox

|mitopedia concept=MiP concept, SUIT protocol, Recommended ::: '''[[SUIT protocol pattern]]:''' 1OctM;2D;3P;4S;5U;6Rot-

|info='''B''' - '''[[SUIT-009]]'''- short protocol for respiration in mitochondrial preparations; mt: |SUIT number=D015_1S;2D;3P;4Rot;5Ama

...ns (isolated mitochondria, permabilized cells, and tissue homogenate)-'''[[SUIT-021]]''' |SUIT number=D035_1GM;2D;2c;3S;4Rot;5Omy;6U;7Ama

|description=[[File:Ce1;1Dig;1S;2D;2c;3P;4Rot;5Ama.png|520px|SUIT-009]] |info='''B''' - '''[[SUIT-009]]''' - short protocol for mt-respiration in ce: non-permeabilized cells

...in the N-,NS- and S- pathway control state on isolated mitochondria- '''[[SUIT-031]]''' |SUIT number=D072_mt;1Q2;1PM;2D;3S;4Rot;5U;6Anox;7Ama

...Pfi;1D;2M.1;3Oct;3c;4M2;5P;6G;7S;8Gp;9U;10Rot;11Ama;12AsTm;13Azd.png|400px|SUIT-RP2]] |info='''A''' -'''[[SUIT-002]]''' - '''[[SUIT reference protocol]] Reference protocol RP2 for permeabilized fibers (pfi)'

::: '''MitoPedia: SUIT''' - '''Substrate-uncoupler-inhibitor titration protocols''' :::<big> » The '''[[SUITbrowser]] helps to find the best SUIT protocol for your research questions'''.</big>

|description=[[File:1GMS;2D-.png|300px|SUIT-018]] ::: '''[[SUIT protocol pattern]]:''' 1GMS;2D;-

...-redox state in the NS- pathway control state on permeabilized cells- '''[[SUIT-031]]''' |SUIT number=D074_ce1;1Dig;1Q2;1PM;2D;3S;4Rot;5U;6Anoxia;7Ama

::: '''[[SUIT protocol pattern]]:'''1S;2D;3P;4Rot- SUIT-009 is a short protocol for simultaneous determination of O₂ flu

|info='''A:''' - '''[[SUIT-003]]''' - [[Coupling-control protocol]] (CCP) in living cells (ce) with ol |SUIT number=D009_ce1;ce2Omy;ce3U;ce4Ama

...le:ce1;1Dig;1PM;2D;2c;3U;4G;5S;(6Oct);7Rot;8Gp;9Ama;10AsTm;11Azd.png|400px|SUIT-RP1 for PBMC and PLT]] |info='''A''' - '''[[SUIT-001]]''' - '''[[SUIT reference protocol]] Reference protocol RP1 for living PBMC and PLT cells (

:::: <big><big><big>'''Development of the MitoFit proficiency test'''</big></big></big> ...n in the field of mitochondrial respiratory physiology. A proficiency ring test is obligatory for certified laboratories.

...on=[[File:Mt;1PM;2D;2c;3U;4G;5S;6Oct;7Rot;8Gp;9Ama;10AsTm;11Azd .png|400px|SUIT-RP1]] |info=[[SUIT-001]] - '''[[SUIT reference protocol]] Reference protocol RP1 for isolated mitochondria, tiss

...File:ce1;1Dig;1PM;2D;2c;3U;4G;5S;6Oct;7Rot;8Gp;9Ama;10AsTm;11Azd.png|600px|SUIT-RP1]] |info='''A''' - '''[[SUIT-001]]''' - '''[[SUIT reference protocol]] Reference protocol RP1 for living cells (ce, ROUTINE r

...'''A''' Linear coupling control with NADH-linked substrates for pce - '''[[SUIT-012]]''' |SUIT number=D052_ce1;1Dig;1PM;2D;2c;3G;4U;5Ama

...s (isolated mitochondria, permeabilized cells, and tissue homogenate)-'''[[SUIT-020]]''' |SUIT number=D032_1PM;2D;2c;3G;4S;5Rot;6Omy;7U;8Ama

...livo Eleonora]]Baglivo E, Timon-Gomez A, Gnaiger E (2023) The cytochrome c test for respirometric quality control of the integrity of the mitochondrial out ...ion by added cytochrome c indicates damage of the mtOM. This respirometric test for mtOM integrity provides a quality control to exclude potential injuries

|abbr=SUIT-catg |description=[[File:SUIT-catg_MitoPathway types.jpg|right|200px]]

...ntrol experiment on which we will add a '''carrier control titration''' to test if it affects our sample or not.

::: '''[[SUIT protocol pattern]]:''' 1PM;2D;2c;3G;4U- ...levant in the presence of [[glutamate-anaplerotic pathway control state]]. SUIT-012 can be extended with the CIV assay module.

...Dig;1D;2M.1;3Oct;3c;4M2;5P;6G;7S;8Gp;9U;10Rot;11Ama;12AsTm;13Azd.png|400px|SUIT-RP2]] |info='''A''' - '''[[SUIT-002]]''' - '''[[SUIT reference protocol]] Reference protocol RP2 for living cells (ce, ROUTINE r

|info='''A''' - '''[[SUIT-002]]''' - '''[[SUIT reference protocol]] Reference protocol RP2 for isolated mitochondria, tiss |SUIT number=D005_mt;1D;2M;3Oct;3c;4M;5P;6G;7S;8Gp;9U;10Rot;11Ama;12AsTm;13Azd

|abbr=CCP-ce S permeability test ...UIT-003]]''' [[Coupling-control protocol]] with succinate for permeability test

...(isolated mitochondria, permeabilized cells, and tissue homogenate)- '''[[SUIT-020]]''' |SUIT number=D033_1PM;2D;3G;4S;5Rot;6Omy;7U;8Ama

|title=Development of a SUIT reference protocol for OXPHOS analysis by high-resolution respirometry. |info=[[MiPNet21.06 SUIT RP]]

...Dig;1D;2M.1;3Oct;3c;4M2;5P;6G;7S;8Gp;9U;10Rot;11Ama;12AsTm;13Azd.png|400px|SUIT-RP2 for PBMC and PLT]] |info='''A''' - '''[[SUIT-002]]''' - '''[[SUIT reference protocol]] Reference protocol RP2 for for living PBMC and PLT cel

|info='''A''' '''[[SUIT-014]]''' |SUIT number=D042_1GM;2D;2c;3P;4S;5U;6Rot;7Ama

::: '''[[Categories of SUIT protocols |SUIT-category]]:''' NS ::: '''[[SUIT protocol pattern]]:''' orthogonal 1PM;2D;3S;4Rot;5U;-

...for isolated mitochondria, tissue homogenate and permeabilized cells- '''[[SUIT-012]]''' |SUIT number=D027_1PM;2D;2c;3G;4U;5Ama

...'''[[SUIT-005]]''' short protocol linked to [[SUIT-002 O2 pfi D006]] - '''SUIT RP2''' (human skeletal muscle) |SUIT number=D011_1OctM;2D;2c;3P;4S;5U;6Rot;7Ama;8AsTm;9Azd

::: '''[[Categories of SUIT protocols |SUIT-category]]:''' NS ::: '''[[SUIT protocol pattern]]:''' orthogonal 1PM;2D;3G;4S;5Rot;6Omy;7U-

...s (isolated mitochondria, tissue homogenate and permeabilized cells) -'''[[SUIT-006]]''' |SUIT number=D034_1PM;2D;3Omy;4U;5Ama

|description=[[File:1PM;2D;2c;3U;4G;5S;6Oct;7Rot;8Gp-.png|400px|SUIT-001]] |mitopedia concept=MiP concept, SUIT protocol, Recommended

|info='''A''' -'''[[SUIT-018]]''' - SUIT protocol for simultaneous determination of O₂ and H₂O |SUIT number=D031_mt;1GMS;2D;3Ama

|description=[[File:1GMS;2D;3Ama.png|290px|SUIT-018]] ...ns (isolated mitochondria, tissue homogenate and permeabilized cells)-'''[[SUIT-018]]'''

::: '''[[SUIT protocol pattern]]:''' 1PM;2D;3G;4S;5U;6Rot- ...se]] and its linked pathways, which could be relevant in some pathologies. SUIT-008 can be easily extended with the CIV assay module.

...IT protocol for mt; mt: isolated mitochondria and tissue homogenate - '''[[SUIT-008]]''' |SUIT number=D026_1PM;2D;2c;3G;4S;4D;5U;6Rot;7Ama;8AsTm;9Azd

::: '''[[SUIT protocol pattern]]:''' 1S;2Rot;3D;4Ama ...ntrations to reach [[OXPHOS]], allows us to harmonize this protocol with [[SUIT-006 O2 mt D022]] for quality control and a full assessment of the coupling

:::: The aim of this workpackage is to test measurement conditions for the newly developed modules and the development ::::* Development of protocols for scientific, pre-clinical and algal biotechnology applications

::: '''[[SUIT protocol pattern]]:''' 1GM;2D;3P;4S;5U;6Rot- ...etermine N-pathway [[Oxidative phosphorylation| ''P'']] capacity. SUIT-014 protocols provide an assessment of the coupling control [[LEAK respiration|''L'']]-[[

::: '''[[SUIT protocol pattern]]:''' 1D;2M;3Oct;4M;5P;6G;7S;8Gp;9U;10Rot- ...pathway control state|SGp]]) can be evaluated by using this SUIT protocol. SUIT-002 can be extended with the CIV assay module.

...than speed''' and seek to publish the definitive work that will stand the test of time ..'' - Quoted from [[Vale 2015 Proc Natl Acad Sci U S A |Vale 2015] :::: '''[[MitoPedia: SUIT]] - first steps towards a data bank of SUIT protocols'''

|info='''A: for cells-permeabilized cells''' [[Lemieux 2017 Sci Rep]] - '''[[SUIT-008]]''' |SUIT number=D025_ce1;1Dig;1PM;2D;2c;3G;4S;5U;6Rot;7Ama;8AsTm;9Azd

...e '''SUITbrowser''' to find the substrate-uncoupler-inhibitor-titration ([[SUIT]]) protocol most suitable for addressing your research questions. ...possible to choose multiple research questions at a time, to find the best protocols that address this combination of problems.

::: '''[[SUIT protocol pattern]]:''' 1GM;2D;3S;4Rot;5Omy;6U;- ...rial membrane potential on isolated mitochondria and tissue homogenate. [[SUIT-021 O2 mt D035]] is a protocol which can be performed without the fluoresce

...s (isolated mitochondria, permeabilized cells, and tissue homogenate)-'''[[SUIT-021]]''' |SUIT number=D036_1GM;2D;3S;4Rot;5Omy;6U;7Ama

|info='''A''' - [[Lemieux 2017 Sci Rep]] - '''[[SUIT-008]]'''- O2 pfi D014 protocol was used a a reference protocol in [[MitoEAG |SUIT number=D014_1PM;2D;2c;3G;4S;4D;5U;6Rot;7Ama;8AsTm;9Azd

...sub> (AmR) in ce: non-permeabilized cells, pce: permeabilized cells -'''[[SUIT-009]]''' |SUIT number=D019_ce1;1Dig;1S;2D;3P;4Rot;5Ama

...preparation protocols (such as [[mitochondrial preparations]]), and [[SUIT protocols]]. === 1. O₂ sensor test ===

...mulated respiration and requires addition of pyr and/or Glut. We primarily test nonhuman primate (japanses macaque) permeabilized muscle fibers.

|description=[[File:1S;2D;3P;4Rot;5Ama.png|400px|SUIT-009]] ...t: isolated mitochondria, tissue homogenate and permeabilized cells - '''[[SUIT-009]]'''short protocol for H₂O₂ (AmR) for isolated mi

::: '''[[SUIT protocol pattern]]:''' 1X;2D;3Omy;4U;5Ama ...e assessed by the [https://bioblast.at/index.php/Oligomycin Omy titration test].

...přivová A (2017) Protocols for high-resolution respirometry experiments to test the activity of electron transfer system of pig brain mitochondria. Indian ...as a biological model for research of mitochondrial respiration using SUIT protocols analogous to those, which were previously published for rodent mitochondria

...e:DoerrierC.JPG|left|90px]] Substrate-uncoupler-inhibitor titration (SUIT) protocols – fundamental principles. ...physiology. In conclusion, an optimal design and comprehension of the SUIT protocols is crucial for a proper evaluation of mitochondrial function in healthy and

|description=[[File:1GM;2D;3S;4U;5Rot-.png|400px|SUIT-011]] ::: '''[[SUIT protocol pattern]]:''' 1GM;2D;2c;3S;4U;5Rot-

...le=[[Image:O2k-Protocols.jpg|right|80px|link=O2k-Protocols|O2k-Protocols]] SUIT reference protocol for OXPHOS analysis by high-resolution respirometry. |abstract=Doerrier C, Sumbalova Z, Gnaiger (2016) SUIT reference protocol for OXPHOS analysis by high-resolution respirometry. Mit

...s (isolated mitochondria, tissue homogenate and permeabilized cells) -'''[[SUIT-006]]''' |SUIT number=D055_1PM;2D;3Cat;4U;5Ama

...2</sub> flux and mitochondrial ATP production in permeabilized cells -'''[[SUIT-006]]''' |SUIT number=D085_ce1;1Dig;1PM;2D;3Cat;4U;5Ama

SUIT reference protocol for comparative mitochondrial physiology. ...physiology. In conclusion, an optimal design and comprehension of the SUIT protocols is crucial for a proper evaluation of mitochondrial function in healthy and

|info='''A''' pfi: permeabilized fibers - '''[[SUIT-011]]''' |SUIT number=D024_1GM;2D;2c;3S;4U;5Rot;6Ama

...Mészáros AT, Gnaiger E (2018) High-Resolution FluoRespirometry and OXPHOS protocols for human cells, permeabilized fibers from small biopsies of muscle, and is ...pdf|O2k-brief]][[Image:O2k-Protocols.jpg|right|80px|link=O2k-Protocols|O2k-Protocols]]

::::» [[Stirrer test]] === Protocols ===

...ture of biomedical and patient specimens with respect to experimental SUIT protocols to be applied. ::::» [[Doerrier 2016 Abstract MitoFit Science Camp 2016]]: Development of a SUIT reference protocol for OXPHOS analysis by high-resolution respirometry.

...sm is associated with apoptosis. Cytochrome ''c'' is applied in [[HRR]] to test the integrity of the [[mitochondrial outer membrane]] ([[cytochrome c contr ...itochondria from mouse. Experiment 2019-02-19 P1-02 ([[SUIT-008_O2_mt_D026|SUIT-008 O2 mt D026]], DatLab 7.4).

...olecules inhibit and may uncouple respiration, which makes it essential to test the optimum concentration. :::* More advanced Excel analysis templates for the respective SUIT protocols to calculate the relative mt-membrane potential values are available on thi

...meter (Oroboros Instruments, Austria). Two harmonized [[SUIT]] protocols [[SUIT-004 O2 pfi D010|RP1]] and [[1OctM;2D;3P;4S;5U;6Rot-|RP2]] [1] modified for ...on was positively associated with the score in Digital Symbol Substitution Test (R=0.388; p=0.038).

...cols published over the years, the aim of my STSM consists in comparing my protocols with those used in the host lab in order to harmonize our procedures and, a ...nity; in particular our aim is the interlaboratory comparisons of specific protocols, a critical issue in results interpretation.

...es, using complex protocols with substrate-uncoupler-inhibitor titrations (SUIT) and monitoring simultaneously different functional parameters in a MultiSe ...citrate synthase. Respiratory coupling control ratios and the cytochrome c test indicated that mitochondria were well coupled and the outer mt-membrane was

:::: '''Standard operating procedures and user requirement document: Protocols, terminology, documentation''' ::::» [[WG1 MitoEAGLE protocols, terminology, documentation |WG1 Action]]

...vice to respirometry experiments with substrate-uncoupler-inhibitor (SUIT) protocols. ...e-uncoupler-inhibitor titration (SUIT) protocols – fundamental principles. SUIT reference protocol: RP1&RP2

:::: 14:45 Start O2k demo experiment - [[SUIT-009 AmR ce-pce D019]] (O₂ and H₂O₂ flux wi ...nstrumental quality control: SOPs from cleaning to instrumental background test

...n and controversial interpretation (Chacko et al 2014). Respirometric SUIT protocols for OXPHOS analysis (Sjoevall et al 2010, 2013, Pecina et al 2014) need to

...ciated with mitochondrial dysfunction. The aim of the present study was to test the hypothesis that oxidative phosphorylation and electron transport capaci [[File:1GM;2D;2c;3S;4U;5Rot;6Ama.jpg|right|300px|link=SUIT-011 |SUIT-011]]

:::: '''Standard operating procedures and user requirement document: Protocols, terminology, documentation''' ::::» [[MitoEAGLE protocols, terminology, documentation |WG1 Project application]]

...verestimated due to a contribution of NADH-linked respiration, F(N) (see [[SUIT-002]]). ::::* {{Template:SUIT M high}}

...Mészáros AT, Gnaiger E (2018) High-Resolution FluoRespirometry and OXPHOS protocols for human cells, permeabilized fibres from small biopsies of muscle, and is == HRR: high-resolution respirometry and SUIT protocols ==

...itrations – a bad combination for an instrumental O₂ background test?'''<br> ...ns affecting the O₂ concentration and thus interfering with the test. Instead of cell culture media, use [[MiR05]], [[MiR06]] or a strongly buff

...month interventions. Mitochondrial respiration was measured using two SUIT protocols in permeabilized muscle fiber bundles. Data were analyzed using a per-proto One hundred participants completed the ''a priori'' defined intervention protocols: (EX, ''N''=44; CON, ''N''=56). Weight loss was similar for both groups (Po

...common cross-linked respiratory states, allowing for harmonization of both protocols. The fluxes were corrected for the contribution by contaminating cells. ...put and maximal lactate accumulation at the end of the ergometric exercise test. In contrast to physical tests, respiration of PBMC and PLT did not differ

...to perform High-Resolution Respirometry using the O2k-FluoRespirometer to test these potential substrates. We expect that mitochondria lacking Slc25a34 wi ...endence only on limited amount of canonical substrates. Furthermore, these protocols can be very helpful for identifying substrates of other mitochondrial metab

...re in the range, for the different parameters assessed. It should serve to test researchers' technical skills and therefore determine if he/she is ready to ...skeletal muscle fibers. The substrates, uncouplers, inhibitors titration (SUIT) protocol that we are assessing is represented in Figure 1. Finally, we wil

...Mészáros AT, Gnaiger E (2018) High-Resolution FluoRespirometry and OXPHOS protocols for human cells, permeabilized fibers from small biopsies of muscle, and is ...f mitochondria-targeted [[SUIT]] (substrate-uncoupler-inhibitor titration) protocols. Measuring respiration of permeabilized cells, allowing direct access to th

...ROS INSTRUMENTS.jpg|right|60px|link=Oroboros Instruments]]Gnaiger E (2010) Test experiments on specifications of the Oroboros Oxygraph. Mitochondr Physiol ...namic corrections in kinetic studies. Results of an extensive experimental test program are presented as the basis of Oroboros O2k-Specifications, which di

...es in ''p''₅₀ reflect inhibition of COX if used as a diagnostic test. ...hway and coupling control on mitochondrial ''p''₅₀. During SUIT protocols, kinetic data for oxygen was obtained with aerobic-anaerobic transitions in

::::* Benefit from the unique features of [[DatLab |DatLab 7]] with [[SUIT protocols]] ...ovation in mitochondrial respirometry: interlaboratory MitoFit Proficiency Test

...' was performed in local anesthesia using Bergström needle technique. SUIT protocols RP1 and RP2 [4] were used to evaluate oxygen consumption rate (pmol s<sup>- :::# Doerrier C, Sumbalova Z, Krumschnabel G, Hiller E, Gnaiger (2016) SUIT reference protocol for OXPHOS analysis by high-resolution respirometry. Mit

...n tissues under physiological or pathological conditions, we established a SUIT protocol for the analysis of oxidative phosphorylation (OXPHOS) and electro # Pesta D, Gnaiger E (2012) High-resolution respirometry. OXPHOS protocols for human cells and permeabilized fibres from small biopsies of human muscl

=== Cytochrome ''c'' test === ...ty. For this reason, the addition of cytochrome ''c'' is routinely used to test the mtOM permeability and to evaluate the quality of the mitochondrial prep

...ate differential diagnosis of platelet pathologies. Including a functional test for platelet activity and verifying its relevance to individual diagnoses w ...LT Protocols (respirometry/Fluorometry): • AmR_SUIT 9_pce: 1S;2D;3P;4Ama • SUIT 7: ce1;ce2P;ce3Omy;ce4U*;ce5Glc;ce6M2;ce7Rot;ce8S;1Dig*;1U*;1c; 2Ama;3AsTm;

...CCP may be extended, when further respiratory states (e.g. cell viability test; CIV assay) are added to the coupling control module consisting of three co ...POS]], [[Gnaiger 2020 BEC MitoPathways]], [[BEC 2020.1]], [[SUIT-003]], [[SUIT-006]]

...vice to respirometry experiments with substrate-uncoupler-inhibitor (SUIT) protocols. ...e-uncoupler-inhibitor titration (SUIT) protocols – fundamental principles. SUIT reference protocol: RP1&RP2

...rformed an in-vivo phosphorus-31 magnetic resonance spectroscopy (31P MRS) test of the quadriceps muscles during dynamic leg-extension exercise [3] and a p ...Mol. Biol.|Pesta D, Gnaiger E (2012) High-resolution respirometry. OXPHOS protocols for human cells and permeabilized fibres from small biopsies of human muscl

...by using different types of Substrate-Uncoupler-Inhibitor-Titration (SUIT) protocols (SUIT1 and SUIT2) where we evaluated the possible deviations between differ ...unlimited titrations and, therefore, the possibility to apply various SUIT protocols designed specifically for a certain project.

...1 M stock solution is used for addition at the beginning of an experiment (SUIT protocol). ::::: To test the need to add these chelators, after addition of MgG, substrates and samp

...s, an elevated oxygen regime is applied, requiring instrumental background test in the same range of elevated oxygen. ...al O2 background]] for experimental details of the instrumental background test.

...er E, Garcia-Roves PM (2019) Inter-laboratory harmonization of respiratory protocols in permeabilized human muscle fibers. https://doi.org/10.26124/mitofit:ea19 ...tofitea19.MiPSchool.0009.pdf Inter-laboratory harmonization of respiratory protocols in permeabilized human muscle fibers]

...rometer|K-Regio project MitoCom Tyrol]], Oroboros Instruments sent two O2k test units to Oregon in May. The newest Series E has 14 instead of 20 kg, which ...ls, based on the latest experience with [[Pesta_2012_Methods_Mol_Biol|SUIT protocols]] developed for functional mitochondrial diagnosis. Catherine trained Austi

...sed as fitness levels increased. Thus, this study is part of a project to test the hypothesis that athletic conditioning results in greater mitochondrial ...fferent incubation temperatures (38, 40, 42, and 44°C) using standard SUIT protocols. Effect of incubation temperature was analyzed using repeated measures one-

...g|80px|link=https://wiki.oroboros.at/index.php/MitoPedia:_SUIT |MitoPedia: SUIT]] The following SUIT protocols are specifically optimized for PBMC and platelets <br>

...easured by Oroboros Oxygraph-2k high-resolution respirometry applying SUIT protocols RP1 and RP2. Oxygen consumption (pmol.s^-1.mg^-1 tissue ...rial fatty acid oxidation was positively associated with short-term memory test score (CogState, ''R''=0.616; ''p''=0.019). Insulin-induced change in resti

::::* Development of an interlaboratory proficiency ring test, ...mitochondrial capacities and create a Library of protocols: reference SUIT protocols, standard experimental media, and detailed instructions for OXPHOS analysis

This page was created to provide a list and details about DatLab protocols for analysis of mitochondrial respiration. | SUIT-001_O2_mt_D001

...ls.jpg|right|80px|link=http://wiki.oroboros.at/index.php/O2k-Protocols|O2k-Protocols contents]]Preparation of permeabilized muscle fibers for diagnosis of mitoc ...njunction with multiple [[substrate-uncoupler-inhibitor titration]] (SUIT) protocols, respirometric studies of human and animal tissue biopsies improve our fund

...tion protocols not only for PBMC but including PLT, applying respirometric protocols specifically developed for diagnostic monitoring. ...amage of PLT by cryopreservation. Further optimization of cryopreservation protocols for blood cells provide new tools for storage and effective use of these ce

...mitochondria. Fairly simple to highly complex, comprehensive experimental protocols can be applied to assess either basal mitochondrial characteristics or obta ...s capacity, residual oxygen consumption, and, by extension of conventional protocols, intactness of cell membranes. In mitochondrial preparations such as isolat

[[File:SUIT-nomenclature.jpg|right|400px|link= MitoPedia: SUIT|SUIT protocols]] === A1: Diagnostic SUIT protocols of mitochondrial fitness ===

...described in the substrate-uncoupler-inhibitor titration (SUIT) protocol [[SUIT-008 O2 pfi D014]] (Fig. 1) [7]. Results of this study will be presented dur [[File:Doerrier_c_Figure_MiP2018.jpg|left|400 px]] Figure 1. [[SUIT-008 O2 pfi D014]] protocol. 25 µM blebbistatin or carrier was added to Buf

...k if their values are in an acceptable range. This approach could serve to test researchers’ technical skills and therefore determine if they are profici ...it following the substrate-uncoupler-inhibitor titration (SUIT) protocol [[SUIT-008_O2_pfi_D014]] represented in Figure1 [1]. Chemicals were provided by ea

::::# Test different USB ports of the PC (this could be caused by a faulty USB port). ...mnSysProtMenu0 already exists' appears when trying to load instrumental DL-protocols using [[DatLab]].

...k if their values are in an acceptable range. This approach could serve to test researchers’ technical skills and therefore determine if they are profici ...7BL/6J mice aged 14-16 weeks. The substrate-uncoupler-inhibitor titration (SUIT) protocol used in the study is represented in Figure 1 [1]. The presentatio

...a media of 5 different assays. Statistical analyses were performed using t-test to demonstrate significant difference between control flies and Pink1 knock ::::#Pesta D, Gnaiger E (2012) High-resolution respirometry: OXPHOS protocols for human cells and permeabilized fibers from small biopsies of human muscl

...ed by a direct measurement within an experimental assay, or by independent test experiments for critical evaluation of the best estimate for a particular ' ...ation of ''Rox'', and for comparison with ''Rox'' measured at the end of a SUIT protocol after titration of substrates and inhibitors of key elements of th

...ed fibers from arm and leg muscle in Inuit versus Caucasians: A functional test of the uncoupling hypothesis in Greenland. Mitochondr Physiol Network 18.08 ...pling hypothesis was based on a substrate-uncoupler-inhibitor titration ([[SUIT]]) protocol using high-resolution respirometry designed to interrogate coup

...human tissue, training with HEK239 cells, quality control and proficiency test must be accomplished a proof of competence,skills and research quality. Thi

...PNet14.06 Instrumental O2 background|Instrumental O₂ background test]] ::::» [[SUIT protocol]]s

...ow. Select your previously evaluated instrumental O₂ background test file and load the parameters (''i.e.'', intercept and slope) in DatLab. ...'': The default [[data recording interval]] is 2 s, applied with most SUIT protocols. The larger the data recording interval, the lower is the time resolution a

...substrate-uncoupler-inhibitor titration (SUIT) protocol. The cytochrome c test (<5% stimulation in healthy controls) indicated outer mt-membrane integrity ...tochondrial physiology and pathology, and for a variety of O2k-MultiSensor protocols applied for functional diagnosis of mitochondrial performance. The fast and

...ing an experiment due to AmR consumption and dilution by titrations in a [[SUIT protocol]]. Check for any potential effects of the experimental AmR concent ...ed with DatLab7.4. For further information, see: [[Instrumental: Browse DL-Protocols and templates]]

...working with "mitochondria people", getting to know the Oxygraph, methods, protocols. I am well certain that soon I will be also a user of the Oroboros instrume ...ome very diligent and hardworking participants - doing the DL7 proficiency test.

...he approach of '''[[cell ergometry]]''' as the state-of-the-art functional test of mitochondrial competence. ''MitoFit'' establishes an inter-university N ::::# [[MitoPedia: SUIT]]

{'''SUIT protocols in mitochondrial research are designed to:''' || SUIT protocols aim to assess function, not to disrupt DNA.

...16:00-17:00 [[Garcia-Souza LF|Luiz Garcia-Souza]]: "Novel cell viability test by high-resolution respirometry" ...ps://www.ncbi.nlm.nih.gov/pubmed/17406588 Frezza ''et al.'' (2007), Nature Protocols]

...ld carry out a test experiment: After reaching anoxia, add oligomycin as a test for the possibility that ATPsynthase acts as a ATPase and thus maintains a :::: When membrane potentials obtained by different protocols but using the same parameters (binding correction factors) during calculati

...whole blood sampling (''e.g.'', K-EDTA, lithium heparin, sodium citrate). Protocols are optimized on the basis of yield, cell fraction purity, intactness of pl :::::# Specific protocols are used for respiration assessments in living and permeabilized. The type

| White adipose tissue mitochondrial respiration protocols - Experiments and Results discussion | Inter-laboratory harmonization of protocols to set a final consensus for the evaluation of the technical skills needed

| '''Cancer cell models to test metabolic intervention strategies''' ...lication of new respiratory substrate-uncoupler-inhibitor-titration (SUIT) protocols are required due to its success in high-resolution respirometry (HRR) and c

|title=[[Image:O2k-Protocols.jpg|right|80px|link=O2k-Protocols]] :>> [[O2k-Protocols]]

| Training in the Oroboros O2k-Laboratory – from SUIT protocols to interpretation and reporting |Harmonization of respirometric protocols comparing isolated mitochondria and permeabilized muscle fibers

:::::: (b) concepts of respirometric SUIT protocols; :::: An entrance test will be the basis for providing grants to students.

|title=[[Image:O2k-Protocols.jpg|right|80px|link=O2k-Protocols|O2k-Protocols]] ::::» [[O2k-Protocols]]

...management network will be a strategic innovation to develop harmonization protocols towards generating a rigorously monitored data repository on mitochondrial ...y is required to implement quality control systems and optimally harmonize protocols across research groups. The primary challenge is to ‘increase value and r

...your dithionite solution has already been checked, perform a leak current test to evaluate which electrical component is generating the out-of-scale signa ...https://wiki.oroboros.at/index.php/High_signal_at_zero_oxygen Leak current test]'''«<br>

...bad electrical connection between these components. The high zero current test helps you to locate the cause. ...itrations – a bad combination for an instrumental O₂ background test?'''<br>

...ism, provides the basis for substrate-uncoupler-inhibitor titration (SUIT) protocols, and updates the terminology consistent with the MitoEAGLE white paper on M ...lect physiological respiratory states in mitochondrial preparations. These protocols ignored the importance of cataplerotic metabolite depletion in the tricarbo

- »'''[[Proficiency test]]'''«<br> ...g|80px|link=https://wiki.oroboros.at/index.php/MitoPedia:_SUIT |MitoPedia: SUIT]]

...g|80px|link=https://wiki.oroboros.at/index.php/MitoPedia:_SUIT |MitoPedia: SUIT]] |''' Not sure which SUIT protocol to apply?'''<br>