MiPNet20.13 Safranin mt-membranepotential

O2k-Fluorometry: HRR and simultaneous determination of mt-membrane potential with safranin. »Bioblast pdf«

» Versions

OROBOROS (2015-06-09) 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.

• 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. »Bioblast pdf«