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| | {{Keywords Membrane potential}} |
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| {{Labeling | | {{Labeling |
| |area=Respiration, Instruments;methods | | |area=Respiration, Instruments;methods |
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| |additional=O2k-Demo, O2k-MultiSensor, MitoFitPublication | | |additional=O2k-Demo, O2k-MultiSensor, MitoFitPublication |
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| :» The [[OROBOROS Mitochondrial Research Laboratory |Oroboros MitoLab]] uses [[Safranin]] O from Sigma (#S2255, 25 g).
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| :» More details: [[Talk:MiPNet20.13 Safranin mt-membranepotential |»Discussion«]].
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| == Safranin chemical background ==
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| :: Several substances typically used in SUIT protocols may influence the fluorescence signal of safranin when injected into the O2k-Chamber (for instance coloured substances such as cytochrome ''c''). These chemicals should be tested for their effect in a background run without biological sample, and the necessary corrections be applied.
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| === Substances with an effect on the fluorescence signal of safranin ===
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| :* ADP (D)
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| :* Cytochrome ''c'' (c)
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| :* Succinate (S)
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| :* Rotenone (Rot)
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| :* Ascorbate (As)
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| :* TMPD (Tm)
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|
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| === Substances without an effect on the fluorescence signal of safranin ===
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| :* Pyruvate (P)
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| :* Malate (M)
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| :* Glutamate (G)
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| :* Digitonin (Dig)
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| :* Oligomycin (Omy)
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| :* FCCP (U)
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| :* Malonic acid (Mna)
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| :* Antimycin A (Ama)
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| :* DMSO
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| :* Ethanol
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| :* H<sub>2</sub>O<sub>2</sub>
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| :* H<sub>2</sub>O
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|
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| == O2k-Fluo LED2-Module / Safranin and TMRM ==
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| The method is easy to use, since the [[Fluorescence-Sensor]] does not get in direct contact with the sample. Transformation of the fluorescence signal of safranin or [[TMRM]] to mtMP [mV] requires a specific method for each preparation and protein content, which has to be kept constant in all samples using this method.
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| References:
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| # Krumschnabel G, Eigentler A, Fasching M, Gnaiger E (2014) Use of safranin for the assessment of mitochondrial membrane potential by high-resolution respirometry and fluorometry. Methods Enzymol 542:163-81. - [[Krumschnabel_2014_Methods_Enzymol|»Bioblast link«]]
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| # Sumbalova Z, Gnaiger E (2015) High-resolution measurement of mitochondrial membrane potential and respiration – comparison of potentiometric and fluorometric methods. Abstract MiP2015. [[Sumbalova_2015_Abstract_MiP2015|»Bioblast link«]]
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Revision as of 08:38, 10 September 2018
O2k-Fluorometry: HRFR and simultaneous determination of mt-membrane potential with safranin.
|
» »Versions
Oroboros (2015-06-15) Mitochondr Physiol Network
Abstract: Krumschnabel G, Eigentler A, Fasching M, Gnaiger E (2015) O2k-Fluorometry: HRFR and simultaneous determination of mt-membrane potential with safranin. Mitochondr Physiol Network 20.13(01):1-5.
Krumschnabel G, Eigentler A, Fasching M, Gnaiger E (2014) Use of safranin for the assessment of mitochondrial membrane potential by high-resolution respirometry and fluorometry. Methods Enzymol 542:163-81. »Bioblast link«
• O2k-Network Lab: AT_Innsbruck_Oroboros
Template:Keywords Membrane potential
Labels: MiParea: Respiration, Instruments;methods
Organism: Mouse
Tissue;cell: Nervous system
Preparation: Homogenate
Coupling state: LEAK, ET
Pathway: S, ROX
HRR: Oxygraph-2k, O2k-Fluorometer, O2k-Protocol
O2k-Demo, O2k-MultiSensor, MitoFitPublication