RÁJECKÝ, Michal, Antonín LOJEK and Milan ČÍŽ. Differentiating between intra- and extracellular chemiluminescence in diluted whole-blood samples. International Journal of Laboratory Hematology. MALDEN: WILEY-BLACKWELL, 2012, vol. 34, No 2, p. 136-142. ISSN 1751-5521. Available from: https://dx.doi.org/10.1111/j.1751-553X.2011.01370.x. |
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@article{1080654, author = {Rájecký, Michal and Lojek, Antonín and Číž, Milan}, article_location = {MALDEN}, article_number = {2}, doi = {http://dx.doi.org/10.1111/j.1751-553X.2011.01370.x}, keywords = {Chemiluminescence; isoluminol; luminol; phagocyte; whole blood}, language = {eng}, issn = {1751-5521}, journal = {International Journal of Laboratory Hematology}, title = {Differentiating between intra- and extracellular chemiluminescence in diluted whole-blood samples}, volume = {34}, year = {2012} }
TY - JOUR ID - 1080654 AU - Rájecký, Michal - Lojek, Antonín - Číž, Milan PY - 2012 TI - Differentiating between intra- and extracellular chemiluminescence in diluted whole-blood samples JF - International Journal of Laboratory Hematology VL - 34 IS - 2 SP - 136-142 EP - 136-142 PB - WILEY-BLACKWELL SN - 17515521 KW - Chemiluminescence KW - isoluminol KW - luminol KW - phagocyte KW - whole blood N2 - Introduction: The differentiation between extra- and intracellular production of reactive oxygen species (ROS) in whole blood was measured by luminol- and isoluminol-enhanced chemiluminescence (CL). Methods: Azide (total CL inhibition), azide + horseradish peroxidase (HRP, restoring extracellular CL), superoxide dismutase + catalase (depleting extracellular ROS) and HRP (enhancing extracellular CL) were used to modulate luminol-and isoluminol-enhanced CL (10(-6)-10(-3) M luminophores) of 125x diluted whole blood which was activated by both calcium ionophore A23187 (Ca-I) and opsonized zymosan particles (OZP) separately. Results: Both activators stimulated intra-and extracellular production of ROS. Luminol-enhanced CL of Ca-I-activated samples detected the intracellular ROS, and with the addition of HRP detected the extracellular CL as well. CL enhanced with isoluminol in concentrations of 10(-4) M or less was mostly extracellular. There was a mixture of intra-and extracellular CL in OZP-activated samples, probably because of the ingestion of luminophore molecules. Conclusion: Measurement of Ca-I-activated CL enhanced with 10(-4) M luminol is recommended for the detection of intracellular ROS. The addition of HRP leads to the detection of overall ROS production while the OZP-activated system with its addition of HRP can only be used to detect overall ROS production. Ca-I-activated CL enhanced with 10(-4) M isoluminol and with addition of HRP is recommended for the detection of extracellular CL. ER -
RÁJECKÝ, Michal, Antonín LOJEK and Milan ČÍŽ. Differentiating between intra- and extracellular chemiluminescence in diluted whole-blood samples. \textit{International Journal of Laboratory Hematology}. MALDEN: WILEY-BLACKWELL, 2012, vol.~34, No~2, p.~136-142. ISSN~1751-5521. Available from: https://dx.doi.org/10.1111/j.1751-553X.2011.01370.x.
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