+ research techniques i (biology 513) staining. + why stain tissue? most tissues are colorless and...

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+ Research Techniques I (Biology 513) Staining

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$Page 1: + Research Techniques I (Biology 513) Staining. + Why stain tissue? Most tissues are colorless and unless one uses diffraction interference contrast microscopy,$

+

Research Techniques I (Biology 513)

Staining

$Page 2: + Research Techniques I (Biology 513) Staining. + Why stain tissue? Most tissues are colorless and unless one uses diffraction interference contrast microscopy,$

+Staining

Why stain tissue?

Most tissues are colorless and unless one uses diffraction interference contrast microscopy, one cannot view the tissue easily

$Page 3: + Research Techniques I (Biology 513) Staining. + Why stain tissue? Most tissues are colorless and unless one uses diffraction interference contrast microscopy,$

+Staining

$Page 4: + Research Techniques I (Biology 513) Staining. + Why stain tissue? Most tissues are colorless and unless one uses diffraction interference contrast microscopy,$

+Types of biological stains

1) Natural dyes

- hematoxylin (from longwood trees)

- stains nuclei blue/purple

2) Mordants

- metallic salts (iron, aluminium, copper) mixed with a dye to enhance staining

- e.g. iron-hematoxylin stains nuclei and myelin

3) Synthetic dyes

- eosin (stains cytoplasmic elements pink/red)

$Page 5: + Research Techniques I (Biology 513) Staining. + Why stain tissue? Most tissues are colorless and unless one uses diffraction interference contrast microscopy,$

+Specialized stains

Zoological tissue: Masson’s trichrome

Even though it has tri- in its name, it contains 4 dyes

Hematoxylin, scarlet-acid fuchsin, phosphomolybdic-phosphotungstic acid, aniline blue

Nuclei stain black, cytoplasm, keratin, muscle fibers stain red, collagen and mucus stain blue or light green

$Page 6: + Research Techniques I (Biology 513) Staining. + Why stain tissue? Most tissues are colorless and unless one uses diffraction interference contrast microscopy,$

+Staining procedure

$Page 7: + Research Techniques I (Biology 513) Staining. + Why stain tissue? Most tissues are colorless and unless one uses diffraction interference contrast microscopy,$

+Staining procedure

4. Counterstain – background material

5. Dehydrate in ascending strengths of alcohol (removal of all water) prepare to coverslip slide

6. Clear (xylene, toluene, histoclear)

7. Cover with mounting medium (Cytoseal 60, Permount) and coverslip clearing agent dissolves mounting medium ensuring an even

coating of medium

$Page 8: + Research Techniques I (Biology 513) Staining. + Why stain tissue? Most tissues are colorless and unless one uses diffraction interference contrast microscopy,$

+Cover slipping procedure

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