Please use this identifier to cite or link to this item: http://doi.org/10.25358/openscience-7580
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dc.contributor.authorManicam, Caroline-
dc.contributor.authorPitz, Susanne-
dc.contributor.authorBrochhausen, Christoph-
dc.contributor.authorGrus, Franz-Hermann-
dc.contributor.authorPfeiffer, Norbert-
dc.contributor.authorGericke, Adrian-
dc.date.accessioned2022-08-19T10:09:42Z-
dc.date.available2022-08-19T10:09:42Z-
dc.date.issued2014
dc.identifier.urihttps://openscience.ub.uni-mainz.de/handle/20.500.12030/7594-
dc.description.abstractPURPOSE: The removal of excessive melanin pigments that obscure ocular tissue morphology is important to address scientific questions and for differential diagnosis of ocular tumours based on histology. Thus, the goal of the present study was to establish an effective and fast melanin bleaching method for paraffin and frozen mouse and human ocular tissues. METHODS: Paraffin-embedded and frozen ocular specimens from mice and human donors were subjected to bleaching employing two methods. The first employed potassium permanganate (KMnO4) with oxalic acid, and the second 10% hydrogen peroxide (H2O2). To determine optimal bleaching conditions, depigmentation was carried out at various incubation times. The effect of diluents used for 10% H2O2 was assessed using phosphate-buffered saline (PBS), and deionized water. Three different slide types and two fixatives, which were ice-cold acetone with 80% methanol, and 4% paraformaldehyde (PFA) were used to determine the optimal conditions for better tissue adherence during bleaching. All tissues were stained in hematoxylin and eosin for histological evaluation. RESULTS: Optimal bleaching was achieved using warm 10% H2O2 diluted in PBS at 65 degrees C for 120 minutes. Chromium-gelatin-coated slides prevented tissue detachment. Adherence of cryosections was also improved with post-fixation using 4% PFA and overnight air-drying at RT after cryosectioning. Tissue morphology was preserved under these conditions. Conversely, tissues bleached in KMnO4/oxalic acid demonstrated poor depigmentation with extensive tissue damage. CONCLUSIONS: Warm dilute H2O2 at 65°C for 120 minutes rapidly and effectively bleached both cryo- and paraffin sections of murine and human ocular tissues.en_GB
dc.description.sponsorshipDFG, Open Access-Publizieren Universität Mainz / Universitätsmedizinde
dc.language.isoengde
dc.rightsCC BY*
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/*
dc.subject.ddc610 Medizinde_DE
dc.subject.ddc610 Medical sciencesen_GB
dc.titleEffective melanin depigmentation of human and murine ocular tissues : an improved method for paraffin and frozen sectionsen_GB
dc.typeZeitschriftenaufsatzde
dc.identifier.doihttp://doi.org/10.25358/openscience-7580-
jgu.type.dinitypearticleen_GB
jgu.type.versionPublished versionde
jgu.type.resourceTextde
jgu.organisation.departmentFB 04 Medizinde
jgu.organisation.number2700-
jgu.organisation.nameJohannes Gutenberg-Universität Mainz-
jgu.rights.accessrightsopenAccess-
jgu.journal.titlePLoS onede
jgu.journal.volume9de
jgu.journal.issue7de
jgu.pages.alternativee102512de
jgu.publisher.year2014-
jgu.publisher.namePLoSde
jgu.publisher.placeLawrence, Kan.de
jgu.publisher.urihttp://dx.doi.org/10.1371/journal.pone.0102512de
jgu.publisher.issn1932-6203de
jgu.organisation.placeMainz-
jgu.identifier.pmid25025426
jgu.subject.ddccode610de
opus.date.modified2018-08-08T09:04:37Z
opus.subject.dfgcode00-000
opus.organisation.stringFB 04: Medizin: Klinikumde_DE
opus.identifier.opusid27370
opus.importsourcepubmed
opus.institute.number0416
opus.metadataonlyfalse
opus.type.contenttypeKeinede_DE
opus.type.contenttypeNoneen_EN
opus.affiliatedPitz, Susanne
opus.affiliatedBrochhausen, Christoph
opus.affiliatedGrus, Franz-Hermann
opus.affiliatedPfeiffer, Norbert
opus.affiliatedGericke, Adrian
jgu.publisher.doi10.1371/journal.pone.0102512de
jgu.organisation.rorhttps://ror.org/023b0x485-
Appears in collections:DFG-OA-Publizieren (2012 - 2017)

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