Difference between revisions of "MiPNet20.13 Safranin mt-membranepotential"

Latest revision as of 11:26, 27 October 2023

MiPNet20.13 Safranin mt-membranepotential

O2k-FluoRespirometry: HRR and simultaneous determination of mt-membrane potential with safranin or TMRM.

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Oroboros (2019-06-24) Mitochondr Physiol Network

Abstract: Krumschnabel G, Fasching M, Gnaiger E (2019) O2k-FluoRespirometry: HRR and simultaneous determination of mt-membrane potential with safranin or TMRM. Mitochondr Physiol Network 20.13(03):1-5.

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» Current O2k-series: NextGen-O2k Series XB and O2k Series J

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• O2k-Network Lab: AT_Innsbruck_Oroboros

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, SUIT-020, SUIT-020 Fluo mt D033, SUIT-006 Fluo mt D034, SUIT-021 Fluo mt D036, SUIT-021, mt-Membrane

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+{{OROBOROS header page name}}
 {{Publication
-|title=[[Image:O2k-Protocols.jpg|right|80px|link=O2k-Protocols|O2k-Protocols]] O2k-Fluorometry: HRFR and simultaneous determination of mt-membrane potential with safranin.
+|title=[[Image:O2k-Protocols.jpg|right|80px|link=O2k-Protocols|O2k-Protocols]] O2k-FluoRespirometry: HRR and simultaneous determination of mt-membrane potential with safranin or TMRM.
-|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]] »[http://www.bioblast.at/index.php/File:MiPNet20.13_Safranin_mt-membranepotential.pdf Versions]
 |authors=Oroboros
-|year=2015-06-15
+|year=2019-06-24
 |journal=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.''' »[http://www.bioblast.at/index.php/File:MiPNet20.13_Safranin_mt-membranepotential.pdf Versions]
+|abstract=Krumschnabel G, Fasching M, Gnaiger E (2019) O2k-FluoRespirometry: HRR and simultaneous determination of mt-membrane potential with [[safranin]] or [[TMRM]]. Mitochondr Physiol Network 20.13(03):1-5.
+{{MiPNet pdf page linking to MitoPedia}}
-[[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=AT_Innsbruck_Oroboros
 }}
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 |pathways=S, ROX
 |instruments=Oxygraph-2k, O2k-Fluorometer, O2k-Protocol
-|additional=O2k-Demo, O2k-MultiSensor, MitoFitPublication
+|additional=O2k-Demo, O2k-MultiSensor, MitoFitPublication, SUIT-020, SUIT-020 Fluo mt D033, SUIT-006 Fluo mt D034, SUIT-021 Fluo mt D036, SUIT-021,mt-Membrane
 }}
-__TOC__
-{{Template:Technical support integrated}}
-:» The [[OROBOROS Mitochondrial Research Laboratory |Oroboros MitoLab]] uses [[Safranin]] O from Sigma (#S2255, 25 g).
-:» More details: [[Talk:MiPNet20.13 Safranin mt-membranepotential |»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
-:* H<sub>2</sub>O<sub>2</sub>
-:* H<sub>2</sub>O
-== 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. -  [[Krumschnabel_2014_Methods_Enzymol|»Bioblast link«]]
-# 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«]]