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Confocal microscopy study of the distribution, content and activity of mitochondria during Paracentrotus lividus development

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Summary In the present paper we applied confocal microscopy andfluorescence technologies for studying the distribution andthe oxidative activity of sea urchin ( Paracentrotus lividus )mitochondria during development, by in vivo incubating eggsand embryos with cell-permeant MitoTracker probes. Wecalculated, by a mathematical model, the intensity values, the variations of intensity, and the variation index of incorporatedfluorochromes. Data demonstrate that mitochondrial massdoes not change during development, whereas mitochondrialrespirationincreases.Inaddition,startingfrom16blastomeresstage, some regions of the embryo contain organelles moreactive in oxygen consumption. Introduction The confocal laser scanning microscope (CLSM) technology,commercially available since 1983, allows the observationof the inner site of living or fixed cells and tissues, and itis able to capture, through computer, serial optical sectionswith a good contrast, creating three-dimensional models.The development of confocal microscopy was accelerated bycomputer, laser and detector technologies, by interferencefilters and fluorochromes, as well as by highly specializedobjectives.CLSMallowsresearcherstocollectaseriesofopticalsectionsand, compared to the wide-field conventional microscopy,offers advantages as a much better control of depth of field andreduction or removal of background noise away from the focalplane.TheCLSMtechnology,basedonfluorescenceemissions,uses probes able to bind to biological macromolecules,