C11-BODIPY(581/591), an oxidation-sensitive fluorescent lipid peroxidation probe: (micro)spectroscopic characterization and validation of methodology.
C11-BODIPY(581/591) is a fluorescent radio-probe for indexing lipid peroxidation and antioxidant efficacy in model membrane systems and living cells, with excellent characteristics: (i) emission in the visible range of the electromagnetic spectrum, with good spectral separation of the nonoxidized (595 nm) and oxidized (520 nm) forms; (ii) has a high quantum yield and because of this, low labeling concentrations can be used, ensuring minimal perturbation of the membrane whilst retaining favorable signal to noise ratios; (iii) has a good photo-stability and displays very few fluorescence artifacts; (iv) is virtually insensitive to environmental changes, i.e., pH or solvent polarity; (v) is lipophilic and as such easily enters membranes; (vi) once oxidized, C11-BODIPY(581/591) remains lipophilic and does not spontaneously leave the lipid bilayer; (vii) C11-BODIPY(581/591) localizes in two distinct pools within the lipid bilayer, a shallow pool at 18 A and a deep pool at < 7.5 A from the center of the bilayer; (viii) is not cytotoxic to rat-1 fibroblasts up to 50 microM; (ix) is sensitive to a variety of oxy-radicals and peroxynitrite, but not to superoxide, nitric oxide, transition metal ions, and hydroperoxides per se; (x) its sensitivity to oxidation is comparable to that of endogenous fatty acyl moieties.
Drummen
Gregor P C
GP
Department of Biochemistry of Lipids, Institute and Graduate School of Biomembranes, Utrecht University, Utrecht, The Netherlands.
van Liebergen
Lydia C M
LC
Op den Kamp
Jos A F
JA
Post
Jan A
JA
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D016477:Artifacts; D001372:Aza Compounds / Q000032:analysis / Q000737:chemistry*; D002250:Carbon Radioisotopes / Q000032:analysis / Q000737:chemistry*; D002412:Cations / Q000494:pharmacology; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line / Q000187:drug effects; D002462:Cell Membrane / Q000378:metabolism; D005227:Fatty Acids / Q000032:analysis / Q000737:chemistry*; D005347:Fibroblasts / Q000187:drug effects; D005456:Fluorescent Dyes / Q000032:analysis / Q000737:chemistry*; D005609:Free Radicals; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007091:Image Processing, Computer-Assisted; D008051:Lipid Bilayers; D015227:Lipid Peroxidation / Q000187:drug effects; D008055:Lipids / Q000302:isolation & purification; D008563:Membrane Lipids / Q000378:metabolism; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D009569:Nitric Oxide / Q000494:pharmacology; D010084:Oxidation-Reduction; D030421:Peroxynitrous Acid / Q000494:pharmacology; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013481:Superoxides / Q000494:pharmacology
D000975:Antioxidants; D001372:Aza Compounds; C463799:BODIPY581; D002250:Carbon Radioisotopes; D002412:Cations; D005227:Fatty Acids; D005456:Fluorescent Dyes; D005609:Free Radicals; D008051:Lipid Bilayers; D008055:Lipids; D008563:Membrane Lipids; D017382:Reactive Oxygen Species; D013481:Superoxides; D030421:Peroxynitrous Acid; D009569:Nitric Oxide; D006861:Hydrogen Peroxide