Difference between revisions of "MiPNet20.13 Safranin mt-membranepotential"
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:» More details: [[Talk:MiPNet19.19 Safranin DataAcquisitionAnalysis |»Discussion«]]. | :» More details: [[Talk:MiPNet19.19 Safranin DataAcquisitionAnalysis |»Discussion«]]. | ||
== Safranin chemical background == | |||
Several substances typically used in SUIT protocols may influence the fluorescence signal from safranin when injected into the O2k-Chamber (for instance colored substances such as cytochrome ''c''). The chemicals used should be tested for this 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 | |||
== Reference == | == Reference == | ||
* 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 |PMID: 24862266]] | * 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 |PMID: 24862266]] |
Revision as of 12:39, 6 June 2015
Safranin data acquisition and analysis. O2k-Fluorometry: HRR and simultaneous determination of mt-membrane potential with safranin. »Bioblast pdf« |
» Versions
OROBOROS (2015-05-27) 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 19.19(01):1-5.
• 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.
HRR: Oxygraph-2k, O2k-Fluorometer, Protocol"Protocol" is not in the list (Oxygraph-2k, TIP2k, O2k-Fluorometer, pH, NO, TPP, Ca, O2k-Spectrophotometer, O2k-Manual, O2k-Protocol, ...) of allowed values for the "Instrument and method" property.
O2k-Demo, O2k-MultiSensor
- » 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 from safranin when injected into the O2k-Chamber (for instance colored substances such as cytochrome c). The chemicals used should be tested for this 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
Reference
- 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. »PMID: 24862266