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Theory of Staining Done

This document discusses various techniques for staining biological specimens to better visualize structures and differentiate between organisms. It describes 5 main types of staining: 1) vital staining of living cells, 2) staining by elective solubility where dyes are more soluble in certain tissues, 3) chemical staining through chemical reactions between dyes and tissues, 4) metallic impregnation using metallic compounds like silver, and 5) staining with dyes either natural or synthetic. It also discusses concepts like metachromasia where dyes change color upon binding tissue, fluorescent staining using fluorescent dyes, progressive vs regressive staining techniques, and differentiation to remove excess stain.

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14 views17 pages

Theory of Staining Done

This document discusses various techniques for staining biological specimens to better visualize structures and differentiate between organisms. It describes 5 main types of staining: 1) vital staining of living cells, 2) staining by elective solubility where dyes are more soluble in certain tissues, 3) chemical staining through chemical reactions between dyes and tissues, 4) metallic impregnation using metallic compounds like silver, and 5) staining with dyes either natural or synthetic. It also discusses concepts like metachromasia where dyes change color upon binding tissue, fluorescent staining using fluorescent dyes, progressive vs regressive staining techniques, and differentiation to remove excess stain.

Uploaded by

mohamedabdulkadir767
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PPTX, PDF, TXT or read online on Scribd
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THEORY OF

STAINING

By Group B
PURPOSE OF STAINING
 To see organism better
 To differentiate one organism from another
 To determine particular structures
TYPES OF STAINING
PROCESS
1. Vital staining: staining of structure in living cells, divided into:
A. Supra vital: living cells stained by dissociation in staining fluid.
B. Intra vital: by injection of the dye into living organism.
2. Staining by elective solubility: this is the mechanism by which stains that
are dissolved in a solvent are more soluble in the tissue component than
they are in the solvent.
E.g.: oil red
Lipid Staining: Oil Red 0 And Hematoxylin
CONTINUE….
3. Staining by chemical: chemical reaction between tissue and colorless
solution to produce a color compound.
The reaction either:
A. True dye: dye bind directly to tissue. E.g: PAS
B. Not true dye: dye react with tissue
constituents and form a colored product. E.g:
perl’s Prussian blue

Pearls Prussian Blue- Hemosiderosis Liver - Iron stain


CONTINUE….
4. Metallic impregnation: metallic compound (ammonical silver)
reduced to metallic state producing black deposit on tissue.
Substances that reduce ammonical silver to metallic silver directly
called argentaffin. E.g: melanin
While substances those need mordant called argyrophil. E.g:
reticulin fiber
silver technique for melanin

Reticulin fiber stained by silver technique


CONTINUE….

5. Staining with dyes: the largest method of staining technique,


according to the nature dyes divided into:
A.Natural dyes: produced from natural source. E.g: hematoxylin
B.Synthetic dyes: a group of organic chemical compounds.
E,g: eosin
CONTINUE….
 Any true dye contain two groups:
A. Chromogen group: group responsible for coloration by absorption of visible
light.
B. Auxochromic group: an ionizing group convert chromogen into true dye which
increase intensity of color.
 The autochrome may be basic:
A. Basic dye: hematoxylin
B. Acidic dye: Eosin
C. Natural dye: Romanowsky dye
METACHROMAISA
 Dyes combine with tissue and give colored compound different from the original
color of dye and color of tissue.
 Dye is known as metachromatic dye, e.g: toluidine and methylene blue.
 Methylene blue and toluidine blue are absorbed by a variety of basophilic
substrate in the tissue. Chromatin stains orthochromatically blue, but cartilage
matrix, mast cell granules metachromatically reddish purple.
FLUORESCENT STAINING
 They have ability of converting ultraviolet light into visible light
when combined with tissue.
 We use fluorescent microscope
 E.g: fluorochrome dye and acridine orange
PROGRESSIVE AND REGRESSIVE STAINING

 Progressive staining: this technique is one in which the different


elements in the tissue are colored in sequence at the correct time.
 E.g: mayer’s hematoxylin
 Regressive staining: this technique is one in which the tissue is first
overstained and then stained or differentiated by removing excess
stain from unwanted parts of the tissue.
 E.g: harris hematoxylin
DIFFERENTIATION
 De-staining or differentiation: is the removal of washing out of the excess
stain until the color is retained only in the tissue components to be
studies.
 Differentiation is used only with regressive stain and can be done by :
 Washing in simple solutions include: water, alcohols
 Acid solutions e.g: picric acid
 Oxidizing agents
DIRECT AND INDIRECT STAINING
 The stains work with out adding mordant is called direct staining
E.g: methylene blue
 While the stains that need mordant is called indirect staining
E.g: hematoxylin
THANKS ANY
QUESTION ?

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