Uncompensated Polychromatic Analysis Of Mitochondrial Membrane Potential Using JC‐1 And Multilaser Excitation
S. de Biasi, L. Gibellini, A. Cossarizza
Published 2015 · Chemistry, Medicine
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The lipophilic cation JC‐1 (5,5′,6,6′‐tetrachloro‐1,1′,3,3′‐tetraethyl‐benzimidazolyl carbocyanine iodide) has been used for more than 20 years as a specific dye for measuring mitochondrial membrane potential (ΔΨm). In this unit, we revise our original protocol (that made use of a single 488 nm laser for the detection of monomers and aggregates, and where compensation was an important step) to use dual‐laser excitation. Moreover, thanks to recently developed multilaser instruments and novel probes for surface and intracellular markers, JC‐1 can be utilized by polychromatic flow cytometry to simultaneously detect, without any compensation between fluorescences, ΔΨm along with other biological parameters, such as apoptosis and the production of reactive oxygen species. © 2015 by John Wiley & Sons, Inc.
This paper references
Source for The Handbook-A Guide to Fluorescent Probes and Labeling Technologies from Invitrogen/Molecular Probes
SPICE: Exploration and analysis of post‐cytometric complex multivariate datasets
M. Roederer (2011)
VDAC phosphorylation, a lipid sensor influencing the cell fate.
Cécile Martel (2014)
Data analysis in flow cytometry: The future just started
E. Lugli (2010)
Reduction in mitochondrial potential constitutes an early irreversible step of programmed lymphocyte death in vivo
N. Zamzami (1995)
Mitochondrial cytochrome c release in apoptosis occurs upstream of DEVD‐specific caspase activation and independently of mitochondrial transmembrane depolarization
E. Bossy-Wetzel (1998)
Multiparametric analysis of cells with different mitochondrial membrane potential during apoptosis by polychromatic flow cytometry
L. Troiano (2007)
Spectral Absorption and Fluorescence of Dyes in the Molecular State
EDWIN E. Jelley (1936)
Mitochondrial functionality and mitochondrial DNA content in lymphocytes of vertically infected human immunodeficiency virus-positive children with highly active antiretroviral therapy-related lipodystrophy.
A. Cossarizza (2002)
Mitochondria are selective targets for the protective effects of heat shock against oxidative injury.
B. Polla (1996)
The Release of Cytochrome c from Mitochondria: A Primary Site for Bcl-2 Regulation of Apoptosis
R. Kluck (1997)
JC‐1, but not DiOC6(3) or rhodamine 123, is a reliable fluorescent probe to assess ΔΨ changes in intact cells: implications for studies on mitochondrial functionality during apoptosis
S. Salvioli (1997)
J-aggregate formation of a carbocyanine as a quantitative fluorescent indicator of membrane potential.
M. Reers (1991)
Mitochondria alterations and dramatic tendency to undergo apoptosis in peripheral blood lymphocytes during acute HIV syndrome
A. Cossarizza (1997)
Honey Bee Hemocyte Profiling by Flow Cytometry
William J. Marringa (2014)
Mitochondrial heterogeneity during staurosporine-induced apoptosis in HL60 cells: analysis at the single cell and single organelle level.
S. Salvioli (2000)
Subject classification obtained by cluster analysis and principal component analysis applied to flow cytometric data
E. Lugli (2007)
The protease inhibitor atazanavir triggers autophagy and mitophagy in human preadipocytes
L. Gibellini (2012)
Protective effect of N-acetylcysteine in tumor necrosis factor-alpha-induced apoptosis in U937 cells: the role of mitochondria.
A. Cossarizza (1995)
Polychromatic flow cytometry: A rapid method for the reduction and analysis of complex multiparameter data
U. Petrausch (2006)
JC-1: alternative excitation wavelengths facilitate mitochondrial membrane potential cytometry
A. Perelman (2012)
Mitochondria and the Autophagy–Inflammation–Cell Death Axis in Organismal Aging
D. Green (2011)
Mitochondrial modifications during rat thymocyte apoptosis: a study at the single cell level.
A. Cossarizza (1994)
Flow Cytometric Analysis of Mitochondrial Membrane Potential Using JC‐1
A. Cossarizza (2001)
Mitochondrial membrane permeabilization in cell death.
G. Kroemer (2007)
Mitochondria: master regulators of danger signalling
L. Galluzzi (2012)
A new method for the cytofluorimetric analysis of mitochondrial membrane potential using the J-aggregate forming lipophilic cation 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolcarbocyanine iodide (JC-1).
A. Cossarizza (1993)
Flavokawain A Induces Apoptosis in MCF-7 and MDA-MB231 and Inhibits the Metastatic Process In Vitro
N. Abu (2014)
Annexin-Phospholipid Interactions. Functional Implications
M. Lizarbe (2013)
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L. Gibellini (2015)
GW9508 ameliorates cognitive impairment via the cAMP-CREB and JNK pathways in APPswe/PS1dE9 mouse model of Alzheimer's disease
Yu-Hang Gong (2020)
Guidelines for the use of flow cytometry and cell sorting in immunological studies.
A. Cossarizza (2017)
Sustained early disruption of mitochondrial function contributes to arsenic-induced prostate tumorigenesis
Bhupendra Pal Singh (2016)
Regulating intracellular ROS signal by a dual pH/reducing-responsive nanogels system promotes tumor cell apoptosis
Kai Wei Dong (2019)
Capsaicin Alleviates the Deteriorative Mitochondrial Function by Upregulating 14-3-3η in Anoxic or Anoxic/Reoxygenated Cardiomyocytes
Y. Qiao (2020)
Mitochondria-targeted therapy rescues development and quality of embryos derived from oocytes matured under oxidative stress conditions: a bovine in vitro model.
W. F. Marei (2019)
Guidelines for the use of flow cytometry and cell sorting in immunological studies (second edition).
A. Cossarizza (2019)
Incubation of human sperm with micelles made from glycerophospholipid mixtures increases sperm motility and resistance to oxidative stress
Gonzalo Ferreira (2018)
Proteomic changes in oocytes after in vitro maturation in lipotoxic conditions are different from those in cumulus cells
W. F. Marei (2019)