Histology Technology

Histology Technology

1. Types of Microscopy

   • Transmission EM: high resolution, 1 nm, 400,000X, electron dense and lucent areas
   • Scanning EM: 3D image, surface views, electrons don't pass though specimen
   • Bright Field: *most focused on
   • Phase-Contrast: Used to enhance structural features. diff type of illumination
   • Nomarski Interence: diff type of optics

2. Types of Stains

   • Hemotoxylin and Eosin (H&E).
     • Hematoxylin is a basic blue dye and binds to acidic neg charge like DNA/RNA Matrix.
     • Eosin: acidic pink dye and binds pos basic charge like mitochondria, cytoplasm, collagen fibers
   • PAS (Periodic Acid Schiff). Stains Glycosaminoglycans which have high sugar/carb content.
   • Wright and Giemsa: Special stain for blood.
     • Pink: RBCs and eosinophil granules
     • Purple: WBC nuclei, basophil granules
     • Blue: Cytoplasm of monocytes, lymphocytes
   • Silver Stain: good to show entire process of neurons.
     • Black: intermediate filaments, Nucleolus
     • White: Nucleus
   • Masson's Tri-chrome stain: special stain for nervous tissue. shows diff b/w synaptic layers, fiber layers, and cellular layer
     • Pinkish/purple: plexiform layer
     • Red: cell layer
   • With phase Contrast: of outer retina
     • Red: cell layer
   • Direct Immunocytochemistry: Tag on the primaryAbs. Show cytoskeleton in diff colors, useful to stain protein dif colors
   • Immunohistochemistry
   • Enzyme Histochemistry: rxn product is insoluble & stained
     • Alkaline-phosphatase staining in kidney tubule
     • Calcium phosphatase. staining for calcium ions
     • Cytochrome Oxidase : of chicken retina

3. Types of Sectioning

   • Cross Section
   • Longitudinal Section
   • En Face:

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