Difference between revisions of "MiPNet20.13 Safranin mt-membranepotential"
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|title=[[Image:O2k-Protocols.jpg|right|80px|link=O2k-Protocols|O2k-Protocols]] O2k-Fluorometry: HRR and simultaneous determination of mt-membrane potential with safranin. | |title=[[Image:O2k-Protocols.jpg|right|80px|link=O2k-Protocols|O2k-Protocols]] O2k-Fluorometry: HRR and simultaneous determination of mt-membrane potential with safranin. | ||
|info=[[File:PDF.jpg|100px|link=http://wiki.oroboros.at/images/5/52/MiPNet20.13_Safranin_mt-membranepotential.pdf |Bioblast pdf]] | |info=[[File:PDF.jpg|100px|link=http://wiki.oroboros.at/images/5/52/MiPNet20.13_Safranin_mt-membranepotential.pdf |Bioblast pdf]] | ||
|authors= | |authors=Oroboros | ||
|year=2015-06-15 | |year=2015-06-15 | ||
|journal=Mitochondr Physiol Network | |journal=Mitochondr Physiol Network | ||
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[[Image:O2k-Publications.jpg|60px|left|link=O2k-Publications:_Topics|O2k-Publications in the MiPMap]] 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«]] | [[Image:O2k-Publications.jpg|60px|left|link=O2k-Publications:_Topics|O2k-Publications in the MiPMap]] 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«]] | ||
|mipnetlab= | |mipnetlab=AT_Innsbruck_Oroboros | ||
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{{Template:Technical support integrated}} | {{Template:Technical support integrated}} | ||
:» The [[OROBOROS Mitochondrial Research Laboratory | | :» The [[OROBOROS Mitochondrial Research Laboratory |Oroboros MitoLab]] uses [[Safranin]] O from Sigma (#S2255, 25 g). | ||
:» More details: [[Talk:MiPNet20.13 Safranin mt-membranepotential |»Discussion«]]. | :» More details: [[Talk:MiPNet20.13 Safranin mt-membranepotential |»Discussion«]]. | ||
Revision as of 10:30, 17 October 2017
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Oroboros (2015-06-15) Mitochondr Physiol Network
Abstract: Krumschnabel G, Eigentler A, Fasching M, Gnaiger E (2015) O2k-Fluorometry: HRR and simultaneous determination of mt-membrane potential with safranin. Mitochondr Physiol Network 20.13(01):1-5. »Versions
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
Labels: MiParea: Respiration, Instruments;methods
Organism: Mouse
Tissue;cell: Nervous system
Preparation: Homogenate
Coupling state: LEAK, ETS"ETS" is not in the list (LEAK, ROUTINE, OXPHOS, ET) of allowed values for the "Coupling states" property.
Pathway: S, ROX
HRR: Oxygraph-2k, O2k-Fluorometer, O2k-Protocol
O2k-Demo, O2k-MultiSensor, MitoFitPublication
MitoPedia O2k and high-resolution respirometry:
O2k-Open Support
- » The Oroboros MitoLab uses Safranin O from Sigma (#S2255, 25 g).
- » More details: »Discussion«.
Safranin chemical background
- 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.
Substances with an effect on the fluorescence signal of safranin
- ADP (D)
- Cytochrome c (c)
- Succinate (S)
- Rotenone (Rot)
- Ascorbate (As)
- TMPD (Tm)
Substances without an effect on the fluorescence signal of safranin
- Pyruvate (P)
- Malate (M)
- Glutamate (G)
- Digitonin (Dig)
- Oligomycin (Omy)
- FCCP (U)
- Malonic acid (Mna)
- Antimycin A (Ama)
- DMSO
- Ethanol
- H2O2
- H2O
O2k-Fluo LED2-Module / Safranin and TMRM
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.
References:
- 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«
- Sumbalova Z, Gnaiger E (2015) High-resolution measurement of mitochondrial membrane potential and respiration – comparison of potentiometric and fluorometric methods. Abstract MiP2015. »Bioblast link«
