Dyes, stains, and special probes in histology

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osmium tetroxide to give a blackcompound; fixation and contrastingof membranes; impregnationaccording to Golgi followed byAgNO 3 solutionSpecial stains:acid and basic dyes(Romanowsky type)GiemsaWrightBluePurplePink-redUsed for blood and bone marrowsmears to demonstrateorthochromatic, polychromatic andmetachromatic propertiesSpecial stains:acid and basic dyes(defined pH)Methylene blueand eosinBluePink (light)Buffered solution of acid and basicdye mixture to demonstratecytoplasmic basophilia on tisuesectionsSpecial stains:acid and basic dyes(polychrome stains)Masson trichromeMallory tripleMovat pentachromeBlueGreenBlue-blackSelective staining of connectivetissue compounds, muscle, fibrinSpecial stains:elastica stainsTaenzer-UnnaWeigertOrceinResorcin fuchsinVerhoeff methodBrown (dark)Blue-blackPurpleBlackElastic tissue, elastic fibrilsSpecial stains:mucin stainsAlcian blueat different pH valuesBlueBlue-greenMucins (glycoconjugates), acid andneutral mucins in gastrointestialepitheliumCombination withother cytochemicalreactions (PAS etc.)MagentaSpecial stains:neurohistologyWeigert hematoxylinMethylene blueCresyl fast violetClassical Nissl orfast Nissl methodsLuxol fast blue(Klüver-Barrera)Violet (nuclei,Nissl bodies)Dark blue (myelinsheaths)Blue-black (myelinsheaths)Deep-blue (nuclei,Nissl bodies)Distinct applications for neurohistology,staining of basophilicstructures by basic dyes, f.e.perikaryon, nuclei, Nissl bodies,glial cells, fibers, myelin sheathsBright blue(myelin sheaths)Special stains:colloidal susp.Trypan blue(vital staining)BlueNontoxic colloidal particles whichdo not label living cells (vitalitymarker of cells in vivo and in vitro,cell suspensions, cultured cells);useful marker of phagocytic cells(cleared by phagocytic system)HistochemistryenzymesSpecific enzymesubstrates forendogenous enzymesChromogendependentSelective cell structure(site of endogenous enzyme)HistochemistryenzymesSpecific probeand defined labels(immunohistology)ChromogendependentCell structure defined by theapplied molecular probe(antigens, antibodies)Fluorochromes:vital staining orstaining of sectionsXanthenesAcridinesTetracycline(cationic, anionicand electroneutralfluorochromes)Different colors(fluorescence)Fluorescence: in vivo labelingof cells and tissue structuresFluorescence: tissue sectionsstained with fluorochromes
Fluorochromes:immunohistologySpecific probeand selected labels(fluorochromes)Different colors(fluorescence)Cell structure defined by theapplied molecular probe(antigens, antibodies etc.)Fluorochromes:in situ hybridizationSpecific probeand selected labels(fluorochromes)Different colors(fluorescence)Cell structure defined by theapplied molecular probe(DNA, RNA)Other labelse.g. enzymes:immunohistology,hybridization andother studiesSpecific probeand selected labels(multiple detectionprinciples)Different colors(chromogenic orfluorescent)Cell structure defined by theapplied molecular probe(antigens, nucleotides etc.)The interaction of dyes with tissuesDyes are obtained either from natural sources or from synthetic production. With theintroduction of the aniline dyes, starting from the mid-19 th centrury, natural dyes have almostlost their role in histology with the exception of hematoxylin from Logwood trees which stillremains its great importance in routine histopathology. P EHRLICH was one of the first tostudy systematically aniline dyes and to introduce scientific methods into the field ofhistological dye staining (EHRLICH P, 1878). Then, GRÜBLER’S Verzeichnis der Farbstoffe(1880) and the Farbstofftabellen (SCHULTZ G and JULIUS P, 1914) gave the first overviews ofthe existing natural and synthetic dyes. Several years later, Colour Index was published as areference guide for manufacturers and consumers by the Society of Dyers and Colourits andthe American Association of Textile Chemists and Colorists (1925). Colour Index is nowpublished as Colour Index International on the web (http://www.colour-index.org/). Colorantsare listed according to Colour Index Generic Names and Colour Index Constitution Numbers.For each product name, Colour Index International lists the manufacturer, the physical form,and the principal uses.Histological staining are based on chemical and physical principles that involve the followingreactions:- electrostatic reactions: dying substances are composed of chromophores (f.e azogroups) and auxochrome groups (f.e. hydroxyl, carboxyl or nitro proups), and thelatter properties define the dye as an acid dye or a basic dye;- metal impregnation: the inherent ability of certain cell structures (argyrophilicelements) to precipitate submicroscopic amounts metal ions (silver ions) which arefurther developed by a reduction process, or the metal impregnation of cellcomponents in a so-called argentaffine reaction following oxidative pretreatments;- selective stainings due to the solubility of the dyes: fat staining with Sudan, Oil redand other fat dyes which relies on differences in solubility of the dye within twomedia, i.e. diffusion from alcoholic dye solution into the fat of specimens;- cytochemical reactions: the application of defined chemical reactions, f.e. chemicalcomplex formation for Fe 3+ staining, enzyme substrate reactions for peroxidases,phosphatases etc., or selective staining of tissue molecules after their chemicalmodification (f.e. introducing reactive groups such as aldehydes for the Feulgen andPAS reaction);- ligand specific probes: the selectivity of cellular staining is defined by the appliedmolecular probe and their specificity for cellular molecules with which they react, f.e.antibodies, lectins, nucleotides.
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