Section 1-11 염색 (Staining - 건양대학교 ?? trypan blue 및 trypan red 수용액과 india ink를 사용 Application of staining 생체염색(vital stain) • 종류 • 초생체 염색(supravital staining)

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  • Section 1-11

    (Staining)

  • (dye) (stain)

    Staining() microscopic image contrast ()

    Stain

    http://en.wikipedia.org/wiki/File:Microscope_with_stained_slide.jpg

  • (benzene)

    Benzene C6H6 , (radical)

    benzene 2 2 () quinone (quinoid compound) .

    Dye ()

  • (chromophores)

    ,

    .

    ethylene (>C=CC=O),

    thiocarbonyl (>C=S), azo (-N=N-), nitro (-N=O), oxy-azo (-N=N-), quinone ring

    Dye ()

  • (chromogen)

    , (chromogen)

    .

    Dye ()

  • (auxochrome)

    2 .

    .

    amino (-NH2), hydroxyl (-OH),

    carboxyl (-COOH), sulfon (-SO3H), amino (-N(R2))

    Dye ()

  • = + +

    Dye ()

  • Leuco compounds ()

    leucobase () .

    PAS Schiff reagent .

    Dye ()

  • (mordants)

    ( )

    lake (dye + mordant)

    hematoxylin alum (NH4, K) , Gram iodine

    chromium, aluminium, iron, sulfate

    Dye ()

  • (accentuators, )

    ,

    lake , .

    methylene blue patassium hydroxide carbol fuchsin phenol

    Dye ()

  • (natural dyes) Hematoxylin

    Mexico Campeche logwood tree

    hematein

    hematein (ripening) .

  • (natural dyes) Hematoxylin

  • (natural dyes) Indigo

    ' , (indogofera)

    Negri (Negri body) Cocchineal (Carmine)

    cocchineal alum

    ,

  • (natural dyes) Orcein

    () Lecanora tinctoria Rocella tinctoria lecanoric acid

    (sex chromatin body) B virus (HBs Ag)

    Saffron Crocus (Saffron )

    alcohol

  • (artifical dyes, synthetic dyes) (acid dyes, anionic dyes)

    ( sodium )

    Na+ - (Chromogen + Auxochrome) - alcohol

    eosin Y, light green, erythrosine, picric acid,

    acid fuchsin OH, COOH, NO2 SO2OH

    , carboxylic acid sulfurous acid sodium .

  • (artifical dyes, synthetic dyes) (basic dyes, cationic dyes)

    ( chloride, sulfate, acetate )

    (Chromogen + Auxochrome) + -Cl- alcohol ,

    NH2, NHCH3, N(CH3)2 NH .

    basic fuchsin, methyl blue, methyl green,

    thionine, toluidine blue

  • (artifical dyes, synthetic dyes) (neutral dyes)

    ( )

    Romanowsky, polychrome methylene blue eosin

    Wright Giemsa , neutral red light green

  • (artifical dyes, synthetic dyes) (amphoteric dye, )

    (+)

    pH ( ) ()

    carmine carminic acid pH 4.5 , (-) , (+)

  • Colour index Number (CIN) Direct Fast Scarlet 4BS , , , S.D.C

    (The Society of Dyers & Colourists. England) A.T.C.C (The American of Textile Chemists & Colourists. U.S.A) (colour index)

    () C.I Name C.I Number .

  • Colour index Number

  • (pore

    size) (> sudan )

    (indifferent dye)

  • (solubility, absortion)

    (lysochrome)

    oil red O propylene glycol

  • (absorption)

    , () ,

    hematoxylin alum aluminium sulfate (+) (-)

    ion (pH) ,

  • , (metachromasia)

    (mucin) toluidine blue

    , .

  • , (metachromasia)

    (sulfated mucopolysaccharide), ,

    (cartilage), (mast cell) , , hyaluronic acid

    methylene blue, thionin, toluidin blue, bismark

    brown, safranin, crystal violet, methyl violet, azure

  • Staining principle - physical

  • ( ), van der waals forces, , (covalent bonding)

    (salt)

    pH (+),

    (-) pH (+)

    pH (-)

  • (metalic impregnation)

    (metal salt) , ,

    reticulum ammonical silver (silvering)

  • binding site

    ,

  • pH ( pH) pH

    , pH .

    .

  • (vital stain) (phagocytosis)

    .

    Application of staining

  • (vital stain)

    (intravital staining)

    trypan blue trypan red india ink

    Application of staining

    http://www.ricercaitaliana.it/images/rif/PRIN/2004065790_002/imm_0003.jpg

  • (vital stain)

    (supravital staining)

    ,

    (cell inclusions)

    methylene blue, Janus green B, neutral red

    Application of staining

    //upload.wikimedia.org/wikipedia/commons/9/99/Reticulocytes_Human_Blood_Supravital_Stain.jpg

  • (simple stain) alcohol

    , methylene blue 0.2% ,

    Application of staining to fixed tissue

  • (direct staining) (mordant)

    congo red, sirius red

    Application of staining to fixed tissue

  • (mordants) (indirect staining)

    , lake --

    hematein, celestine blue B(mordant blue 14), alizaline red S(mordant red 13)

    Application of staining to fixed tissue

  • (salts of heavy metals) (AgNO3) ,

    (silver ion, Ag+)

    (silver impregnation)

    (argyrophilic reaction) (argentaffin reaction) (induced argentaffin

    reaction)

    Application of staining to fixed tissue

  • (argyrophilic reaction)

    , developer

    Helicobacter pylori

    Application of staining to fixed tissue

    http://www.oculist.net/downaton502/prof/ebook/duanes/pages/v7/ch031/013f.htmlhttp://vet.sagepub.com/content/46/6/1181/F2.expansion.html

  • (argentaffin reaction)

    developer .

    Application of staining to fixed tissue

    http://www.americanmastertech.com/store/main.aspx?p=ItemDetailStyles&item=KTFMA

  • (induced argentaffin

    reaction)

    ( aldehyde )

    Application of staining to fixed tissue

  • van Gieson

    picric acid acid fuchsin

    Application of staining to fixed tissue

    http://eulep.pdn.cam.ac.uk/pathbase2/Search_Pathbase/image.php?image_number=3411.0

  • (progressive staining)

    ,

    Mayer's hematoxylin , Gills hematoxylin

    Progression processing

  • (progressive staining)

    Progression processing

  • (regressive staining)

    ( ) (overstaining) ()

    () .

    Harris' hematoxylin

    Progression processing

  • (regressive staining)

    Progression processing

  • ( : differentiation, decoloration)

    ( ) ( ) .

    , , ,

    Progression processing

  • ( : differentiation, decoloration) ,

    alum hematoxylin acid alcohol(1%

    HCl / 70% alcohol)

    eosin 0.1% ~ 0.5% ammonia / 70%

    alcohol

    alcohol , alcohol

    Progression processing

  • ( : differentiation, decoloration) ,

    pH ,

    , pH .

    pH 1.5~2.0 hematoxylin pH 2.2~2.9

    pH 5.5~6.0 eosin pH 4.5

    Progression processing

  • ( : differentiation, decoloration)

    , , dye-mordant (-)

    Verhoeff's elastic fiber , hematoxylin ferric chloride

    Progression processing

  • ( : differentiation, decoloration)

    pH 6.0~6.5 wrights

    pH methylene blue eosin pH methylene blue , eosin

    alcohol pH

    ferricyanide, bisulfate, permanganate

    Progression processing

  • (orthochromasia)

    (metachromasia)

    (polychromasia)

    Wright

    Types of chromasia

  • Routine staining ()

  • hematoxylin & eosin

    Wright , Gram , Papanicolaou

    Routine staining

  • Hematoxylin Wihelm von Waldeyer

    (1862~3), H & E

    Hematoxylin , , Hematoxylin campechianum logwood

    Hematoxylin , alcohol ether .

    Routine staining

  • Hematoxylin hematein(C16H12O6) ,

    hematoxylin hematein aluminium

    hematoxylin hematein , (ripening)

    Hematoxylin (ex> , , , ) .

    Routine staining