From Bioblast

high-resolution terminology - matching measurements at high-resolution



Ce1;ce2(Omy);ce3U-.png Ce5S;1Dig;1c-.png

Abbreviation: CCP-ce

Reference: A: Coupling-control protocol, living cells Β»MiPNet08.09 CellRespiration Bioblast pdf

SUIT protocol pattern: 1ce;2ceOmy;3ceU;ce*, or ce1;ce2Omy;ce3U;ce4Rot;ce5S;1Dig;1c;2Ama

The protocols recommended in the SUIT-003 have been designed to study the coupling control state of living cells. In order to study the LEAK respiration, the phosphorylation system needs to be inhibited, which is achieved in this protocol using oligomycin. However, the oligomycin concentration has to be carefully adjusted for different samples and the inhibitory effect on the electron transfer system has to be studied to avoid an underestimation of the ET capacity. SUIT-003 follows a modular approach and can be easily used in combination with the permeability test and/or the assessment of the complex IV capacity.

Communicated by Cardoso LHD, Doerrier C, Huete-Ortega M, Gnaiger E (last update 2021-03-23)

Specific SUIT protocols

SUIT-003 O2 ce D009

1ce;2ceOmy;3ceU-.jpg D009 Traces.png

SUIT-003 O2 ce-pce D020

Ce1;ce1P;ce2Omy;ce3U;ce4Rot;ce5S;1Dig;1c;2Ama;3AsTm;4Azd.png D020 O2 traces.png

SUIT-003 O2 ce D037 and SUIT-003 O2 ce D038

Ce1;ce1Glc;ce2(Omy);ce3U;ce4Ama.png D037 O2 traces.png

Ce1;ce2(Omy);ce3U;ce3Glc;ce3'U;ce4Ama.png D038 O2 trace.png

SUIT-003 O2 ce D039

Ce1;(ce2Omy);ce3U;ce4Rot;ce5Ama.jpg D009 Traces algae.png

SUIT-003 O2 ce D050

Ce1;ce1Snv;(ce2Omy);ce3U;ce4Rot;ce5Ama.png D050 O2 traces.png

SUIT-003 pH ce D067

SUIT-003 O2 ce D067 diagram.png D067 O2 trace.pngD067 pH trace.png

  • SUIT-003 pH ce D067 to study glycolysis with simultaneous measurement of respiration and proton flux, besides coupling control and Crabtree effect.
MitoPedia: SUIT

Steps and respiratory states


Step State Pathway Q-junction Comment - Events (E) and Marks (M)

ce1 ROUTINE ce1
  • ROUTINE respiration in the physiological coupling state R. Externally added permeable substrates could contribute to this respiratory state.
ce2(Omy) L(Omy) ce1;ce2Omy
  • Non-phosphorylating resting state (LEAK state); LEAK-respiration, L(Omy), after blocking the ATP synthase with oligomycin.
  • The addition of oligomycin or its carrier (EtOH) is optional depending on the experimental question to resolve. If we add only the carrier we do not reach LEAK state and we will evaluate if the EtOH alters the ROUTINE state.
ce3U E ce1;ce2Omy;ce3U
ce* ROX ce1;ce2Omy;ce3U;ce*
  • Rox is the residual oxygen consumption in the ROX state, due to oxidative side reactions, estimated either after inhibition of CIII (e.g. antimycin A, myxothiazol), CIV (e.g. Cyanide) or in the absence of endogenous fuel-substrates. Rox is subtracted from oxygen flux as a baseline for all respiratory states, to obtain mitochondrial respiration.


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Strengths and limitations

  • The SUIT-003 is a cell coupling-control protocol. The total cell count (Nce) is the sum of viable cells (Nvce) and dead cells (Ndce).
  • This protocol can be extended with the cell viability test module.
  • For determination of respirometric viability index, it is assumed that both viable and dead cells contain functional mitochondria.
  • When the cell viability test module is not incorporated to the experiment, complex III inhibitors should be used to reach Rox instead of rotenone.
  • When MiR05 is used, the addition of pyruvate is recommended to provide an external fuel substrate.
+ Respirometric cell viability test correlates with other cell viability quantification methods.
+ With the extended module (SUIT-003 O2 ce-pce D020), S-ET capacity can be assessed after the performance of the coupling-control protocol, as well as outer mt-membrane integrity with addition of cytochrome c.
+ CIV activity can also be measured after the cell viability test module.
+ Flexibility of experiment duration: from 1 hour (cell coupling-control protocol) to 2 and a half hours (cell coupling control, cell viability test module and CIV module).
+ Reasonable duration of the experiment.
- Cells with a cell membrane succinate transporter are not suitable for the viability test.
- Pathological profiles could hinder the cell viability test.
- Careful washing is required after the experiment to avoid carry-over of inhibitors and uncoupler.

Compare SUIT protocols


MiPNet08.09 CellRespiration2016-08-11
O2k-Procedures contents
High-resolution respirometry and coupling-control protocol with living cells: ROUTINE, LEAK, ET-pathway, ROX.
HumanBlood cells
Gnaiger 2008 POS2008Gnaiger E (2008) Polarographic oxygen sensors, the oxygraph and high-resolution respirometry to assess mitochondrial function. In: Mitochondrial dysfunction in drug-induced toxicity (Dykens JA, Will Y, eds) John Wiley & Sons, Inc, Hoboken, NJ:327-52.MouseBlood cells
Huetter 2004 Biochem J2004HΓΌtter E, Renner K, Pfister G, StΓΆckl P, Jansen-DΓΌrr P, Gnaiger E (2004) Senescence-associated changes in respiration and oxidative phosphorylation in primary human fibroblasts.

MitoPedia concepts: MiP concept, SUIT protocol, Recommended 

MitoPedia methods: Respirometry 

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