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Optical probes for detection and quantification of neutrophils’ oxidative burst. A review
Authors:Marisa Freitas
Institution:REQUIMTE, Departamento de Química-Física, Faculdade de Farmácia, Universidade do Porto, Rua Aníbal Cunha 164, 4099-030 Porto, Portugal
Abstract:Neutrophils, also known as polymorphonuclear leukocytes (PMN), are the most common type of white blood cells, comprising about 50-70% of all white blood cells. In the event of inflammatory processes, neutrophils display increased mobility, tissue influx ability, prolonged life span, and an increased phagocytic capacity, constituting the initial participants in the cellular defense of the organism. One of the most important defense systems of neutrophils corresponds to their ability to mediate a strong oxidative burst through the formation of reactive oxygen species (ROS) and reactive nitrogen species (RNS). While oxidative burst is important for the elimination of invading microorganisms, the overproduction of ROS and RNS or the impairment of endogenous antioxidant defenses may result to detrimental effects to the host. The nature and the extent of ROS and RNS production by neutrophils in response to different stimuli is, consequently, a matter of extensive research, with scientific reports showing an enormous variability on the detection methodologies employed. This review attempts to provide a critical assessment of the most common approaches to identify and quantify reactive species formed during the neutrophils’ oxidative burst. The detection mechanisms and performance, as well as advantages and limitations of the different methodologies, are scrutinized, focusing on the use of fluorimetric, chemiluminometric and colorimetric probes.
Keywords:APF  2-[6-(4&prime  -amino)-phenoxy-3H-xanthen-3-on-9-yl] benzoic acid  CGD  chronic granulomatous disease  cNOS  nitric oxide synthase constitutive  DAF-FM  4-amino-5-methylamino-2&prime    7&prime  -difluorofluorescein  DAF-FM DA  4-amino-5-methylamino-2&prime    7&prime  -difluorofluorescein diacetate  DAF-FMT  4-amino-5-methylamino-2&prime    7&prime  -difluorofluorescein triazole  DAF-2  4  5-diaminofluorescein  DAF-2 DA  4  5-diaminofluorescein diacetate  DAF-2T  4  5-diaminofluorescein triazole  DCF  2&prime    7&prime  -dichlorofluorescein  DCFH  2&prime    7&prime  -dichlorodihydrofluorescein  DCFH-DA  2&prime    7&prime  -dichlorodihydrofluorescein diacetate  DHR  dihydrorhodamine 123  DPA  10-diphenylanthracene  DPI  diphenyleneiodonium chloride  DTNB  5  5&prime  -dithio-bis-2-nitrobenzoic acid  E+  ethidium  eNOS  nitric oxide synthase expressed in endothelial cells  fMLP  N-formyl-methionyl-leucyl-phenylalanine  GFP  green fluorescent protein E  coli  H2O2  hydrogen peroxide  HbO2  Oxyhemoglobin  HE  hydroethidine or dihydroethidium  HOradical dotels-cdn  " target="_blank">com/sd/entities/rad" class="glyphImg">  hydroxyl radical  HOCl  hypochlorous acid  HPF  2-[6-(4&prime  -Hydroxy)phenoxy-3H-xanthen-3-on-9-yl] benzoic acid  HRP  horseradish peroxidase  HSA  human serum albumin  iNOS  nitric oxide synthase inducible  L-012  8-amino-5-chloro-7-phenylpyrido[3  4-d]pyridazine-1  4-(2H  3H)dione  l-NAME" target="_blank">l-NAME  l-arginine methyl ester" target="_blank">N-nitro-l-arginine methyl ester  MbO2  oxymyoglobin  MCD  monochlorodimedon  MCLA  2-methyl-6[p-methoxyphenyl]-3  7-dihydroimidazo[1  2-α]pyrazin3-one  MetHb  methemoglobin  MetMb  metmyoglobin  MPO  myeloperoxidase  MTT  3-(4  5-dimethylthiazol-2-yl)-2  5-diphenyltetrazolium bromide  NBT  nitroblue tetrazolium  nNOS  nitric oxide synthase expressed in neuronal cells  radical dotels-cdn  NO" target="_blank">com/sd/entities/rad" class="glyphImg">NO  nitric oxide  radical dotels-cdn  NO2" target="_blank">com/sd/entities/rad" class="glyphImg">NO2  nitrogen dioxide radical  N2O3  dinitrogen trioxide  NOS  nitric oxide synthase  1O2  singlet oxygen  O2radical dotels-cdn  &minus" target="_blank">com/sd/entities/rad" class="glyphImg">&minus    superoxide radical  ONOO&minus    peroxynitrite anion  ONOOCO2&minus    nitrosoperoxycarbonate anion  ONOOH  peroxynitrous acid  phox  phagocyte oxidase  PKC  protein kinase C  PMA  phorbol 12-myristate 13-acetate  PMN  polymorphonuclear leukocyte  RNS  reactive nitrogen species  ROOradical dotels-cdn  " target="_blank">com/sd/entities/rad" class="glyphImg">  peroxyl radical  ROS  reactive oxygen species  SNAPF  sulfonaphthoaminophenyl fluorescein  SOD  superoxide dismutase  TMB  3  3&prime    5  5&prime  -tetramethylbenzidine  TNB  2-nitro-5-thiobenzoic acid  WST-1  2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2  4-disulfophenyl)-2H-tetrazolium
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