Overview of staining methods in histology STAINING Different cell or tissue structures are not apparent without staining. Cellular structures exhibit different affinity to staining dyes: alkaline dyes – react with anionic structures BASOPHILIA (basophilic structures in the cell – nucleus, nucleolus, ribosomes, cytoplasm with rough ER) acid dyes – react with cationic groups ACIDOPHILIA – (acidophilic structures in the cell – cytoplasm, smooth ER) no or weak reaction NEUTROPHILIA ROUTINE STAINING with HEMATOXYLINE – EOSIN (HE) Hematoxyline – basic (nuclear) dye Eosin – acid (cytoplasmic) dye • Staining procedure: • paraffin must be removed (dissolved) by xylene • sections are rehydrated in descending series of ethanol (100% 96% 80%) • staining with hematoxyline • differentiation in acid ethanol and water (excess of dye is removed) • staining with eosin • rinsing in water (excess of dye is removed) • dehydration in graded ethanol series (80% 96% 100%) • clearing in xylene Xylen I XylenII 100% 96% H2O hematoxyline acid ethanol ethanol ethanol Xylen IV xylen III 100% 96% H2O eosin H2O ethanol ethanol HEMATOXYLINE – EOSIN (HE) Deparaffination Rehydration Washing Staining Differentiation Clearing Dehydration Washing Staining Washing Automatic slide stainer staining set of boxes with media Hematoxyline Eosin (HE) Results of staining: cell nuclei – blue/violet cytoplasm – pink collagen fibers – pink muscle cells – dark pink/red 2 – Apex linguae (HE) Hematoxyline Eosin Saffron (HES) Results of staining: cell nuclei – blue/violet cell cytoplasm – pink collagen fibers – yellow yellow Masson trichrom 11 – Oesophagus (HES) Azokarmin Aniline blue Orange G (AZAN) Results of staining: cell nuclei – purple cell cytoplasm – pink collagen fibers – blue erythrocytes – orange blue Masson trichrom 99 – umbilical cord (AZAN) Impregnation with AgNO3 68 – Lien (impregnation) Special staining methods Slide 28 – elastic cartilage Staining – orcein Result – red-brown elastic fibers Slide 31 – renal cortex Staining – Weigert-van Giesson Result – cherry-red collagen fibers Special staining methods Slide 49 – vagina - glycogen Staining – Best carmine Result – darc pink glycogen Special staining methods Slide 65 – myocardium Staining – Heidenhain Result – black cardiomyocytes Special staining methods Slide 76 –cortex cerebri Staining – impregnation Result – brown cells Special staining methods Slide 68 – lien Staining – impregnation Result – black reticular fibers Special staining methods Slide 78 – cerebellum Staining – Nissl Result – blue RER, N, ncl. Special staining methods Slide 95 – bone Staining – Schmorl Result – rusty brown bone tissue Special staining methods Slide 87 – nerve - myelin Staining – luxol blue Result – blue myelin Special staining methods Overview of staining methods in histology 2 – Apex linguae (HE) 11 – Oesophagus (HEŠ) 99 – Pupečník (AZAN) 68 – Lien (impregnace) 64, 65 – Myokard (HE, Heidenhain) Slides: Numbers by the slides indicate their position in sets in Microscopic Hall, not in online atlases. They allow you to find them easily and study them using a microscope when the normal classes are opened.