Basic Histo Techniques

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Fixation

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treatment of tissue with chemical agents (fixatives);
preserves tissues

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Fixatives

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coagulate proteins and make them insoluble

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Dehydration

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removal of water because embedding agent is immiscible with water

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Clearing

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treating tissue with agent miscible with melted paraffin eg xylene. Clearing makes tissue transparent

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Embedding

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making a mold; use of paraffin

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Infiltration

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completely immersing the tissue in melted paraffin

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Sectioning

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trimming excess paraffin, or any embedding material, used from tissue- 5micrometers

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Mounting

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placing sections on slides; could be on adhesive-coated or stained slides

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H&E

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Hematoxylin and Eosin

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hematoxylin

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Hematoxylin: Basic stain that preferentially colors acidic components of the cell e.g basophilic components: nucleus; regions rich in ribosomes

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Eosin

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Acidic dye that stains the compponents of a cell pinkish color. Cytoplasmic components with basic pH stain pink- acidophilic

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Toluidine blue (Tol B1)

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its molecules polymerize with each other when exposed to high concentrations of polyanions in tissue

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Metachromatic

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A tissue or component that stains purple (rich in polyanions)

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Metachromasia

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Toluidine blue exhibits this charateristic: blue stain

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masson's trichome

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Nuclei=dark blue
Muscle, keratin, cytoplasm= red
Mucinogen, collagen= light blue

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Staining

Back 16

trimming excess paraffin, or any embedding material, used from tissue- 5micrometers
treating tissues with reagent or dye that makes certain structures visible without affecting others

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cryotome

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rotary microtome in a frozen section environment

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Microtome

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An instrument used to cut tissue into thin sections for microscpic examination

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Orcein's elastic stain

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elastic fibers: brown

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Weigert's elastic stain

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Elastic fibers: blue

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Methamine Silver stain

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reticular fibers: black, and in polarized light using red plate, crystals they look yellow

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Iron hematoxylin (FeH)

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muscle striations, nuclei, RBCs: black

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Periodic acid-Schiff (PAS)

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Glycogen and carb rich molecules: magenta

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Wright's and Geimsa stains (blood cell staining)

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RBS, eosinophil granules:pink
cytoplasm of monocytes, lymphocytes: blue

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Lipochrome

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intermediate of lipid breakdown

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FITC

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Fluorescein Isothicyanate: used as a fluorescent label for proteins e.g antibodies

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Hemosiderin

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Insoluble protein that contains iron
especially found in liver

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HRP

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Horseradish peroxidase;more stable, more popular than AP;
increases detectability of target molecule

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AP

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alkaline phosphatase; dephosphorylates many molecules

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Direct immunocytochemistry

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antibody labelling; Ab labelled with fluorescent dye, then permitted to react with macromolecule

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Costamere

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structural component of striated muscle cells; connects sarcomere of muscle to cell membrane

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Resolution

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the ability to show that two distinct objects are separated by a distance

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Histochemistry

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method of tissue staining that provides information anout the presence and location of intracellular and extracellular macromolecules

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Autoradiography/radioautography

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method used for localizing and investigating a specific temporal sequence of events.
uses incorporation of radioactive isotopes into macromolecules, then visualized by the use of an overlay of film emulsion

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Indirect Immunocytochemistry

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Ab prep against 1o Ab. 1o Ab binds antigen, wash unbound, then labelled Ab is alloed to bind complex= 2o complex. viewed via fluorescent microscopy

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Immunocytochemistry

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ferritin (electron dense molecule) labeling may be used for doth direct and indirect

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Confocal microscopy

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treat samples with fluorescent dye; laser beam thru specimen causees light emission from dyes; single pts gathered y photomultiplier tube; 3D image produced

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Electron microscopy

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focusing a beam of electrons; resolution of 0.005nm