Please use this identifier to cite or link to this item: http://repositorio.unicamp.br/jspui/handle/REPOSIP/200344
Type: Artigo de periódico
Title: Safranine As A Fluorescent Probe For The Evaluation Of Mitochondrial Membrane Potential In Isolated Organelles And Permeabilized Cells.
Author: Figueira, Tiago R
Melo, Daniela R
Vercesi, Aníbal E
Castilho, Roger F
Abstract: The mitochondrial electrical membrane potential (Δψ) is the main component of the proton motive force (Δp) generated across the inner mitochondrial membrane during electron flow through the respiratory chain. Among the techniques available to assess Δψ, methods that rely on the spectrophotofluorometric responses of dyes are widely employed for whole suspensions of isolated mitochondria or permeabilized cells. Safranine is one of the dyes currently used most often for this purpose. Safranine is a lipophilic cationic dye that undergoes optical shifts upon its potential-dependent distribution between the external medium and the intramitochondrial compartment and on its stacking to inner mitochondrial membrane anionic sites. The association between the optical changes of safranine and the membrane potential allows unknown Δψ values to be estimated from an equation describing their relationship. Here, we describe the use of safranine as a fluorescent indicator of Δψ in isolated mitochondria and digitonin-permeabilized cells. We present suitable conditions to employ safranine as a Δψ indicator.
Subject: Animals
Digitonin
Female
Fluorescent Dyes
Indicators And Reagents
Membrane Potential, Mitochondrial
Mitochondria, Liver
Mitochondrial Proteins
Mitochondrial Size
Organelles
Pc12 Cells
Permeability
Phenazines
Rats
Rats, Wistar
Citation: Methods In Molecular Biology (clifton, N.j.). v. 810, p. 103-17, 2012.
Rights: fechado
Identifier DOI: 10.1007/978-1-61779-382-0_7
Address: http://www.ncbi.nlm.nih.gov/pubmed/22057563
Date Issue: 2012
Appears in Collections:Unicamp - Artigos e Outros Documentos

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