STAINING Flashcards by Toni Mae Inductivo

● Derived from the heartwood of the mexican tree

Hematoxylin

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Hematoxylin campechianum

Hematoxylin

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the active coloring substance

Hematein

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If hematoxylin is not ripened,
it _________ be used.

Cannot

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It was previously used in the textile industry

Hematoxylin

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Hematoxylin: First used by __________________________ in 1863 for staining tissue samples.

Wilhelm von Waldeyer-Hartz

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● Derived from the female Coccus cacti bug
● A natural dye

Cochineal Dyes

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Cochineal dye + alum =

carmine stain

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○ a chromatin stain
○ may be mixed with other
material to create another
carmine stain (derivatives)

Carmine

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Used for HBsAg demonstration

Orcein

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STAINING: NATURAL DYES (4)

Hematoxylin
Cochineal Dyes
Orcein
Saffron

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are obtained from plants and
animals

Natural dyes

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also means oxidation

Ripening

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● Made up of:
○ carmine + picric acid

Picro Carmine

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● Used for neuropathological studies

Picro Carmine

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● Made up of:
○ carmine + aluminum HYDROXIDE

Mucicarmine

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● Used for the demonstration of
Cryptococcus neoformans and
mucin

Mucicarmine

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● Made up of:
○ carmine + aluminum CHLORIDE

Best Carmine

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Also known as “Coal Tar Dyes”

SYNTHETIC DYES

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Derived from benzene and collectively known as

“Aniline Dyes”

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Used for glycogen demonstration

Best Carmine

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CHROMOPHORES (4)

● Quinoid Ring
● Azo Groups
● Xanthene
● Quinone-Imine Group
○ Oxazin
○ Thiazins

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Derived from benzene and collectively known as “Aniline Dyes”

SYNTHETIC DYES

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AUXOCHROMES (2)

● Cationic Auxochrome
● Anionic Auxochrome

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AUXOCHROMES: Amino Group

Cationic Auxochrome

AUXOCHROMES: Hydroxyl And Carboxyl Groups

Anionic Auxochrome

DYE MODIFIERS ● Attached on benzene ring (3)

○ Ethyl groups ○ Methyl groups ○ Sulphonic Acid

● Majority of tissue-dye reactions ● Examples: ○ Neutral Red ○ Light Green

Electrostatic

● Examples: ○ Congo Red ○ Carmine ○ Weigert-type resorcinol dye

Hydrogen bonding

● Examples: ○ Alum Hematoxylin Solutions

Van der Waals Forces

● Examples: ○ Sudan dyes ● SUDANOPHILIA

Physical Staining

property of tissues to be stained with fat or oil-soluble dyes, regardless of the type of dye used, due to their essential lipid nature

SUDANOPHILIA

● Examples: ○ Janus Green

Natural Affinity

Benzene + Chromophore =

Chromogen

Chromogen + Auxochrome =

Dye

Responsible for "Coloring Property"

Chromophore

Responsible for "Dyeing Property"

Auxochrome

Imparts color Temporarily

Chromogen

● A benzene ring with an attached chromophore. ● Impart color temporarily

Chromogen

Imparts color ALMOST permanently

Dye

● Impart color ALMOST permanently ○ Strong decolorizer may remove the color

Dye

Which part of the synthetic dye is responsible for coloring property?

Chromophore

Which part of the synthetic dye is responsible for dyeing property?

Auxochrome

METHODS OF STAINING (4 GROUPS)

● According to the presence of a mordant ● According to the presence of a differentiator ● According to the resultant color of the tissue ● Vital Staining

serves as a link or a bridge between the stain and the tissues.

Mordant

Also known as decolorizer

Differentiator

According to the resultant color of the tissue (2)

○ Orthochromatic staining ○ Metachromatic staining

ACCORDING TO THE PRESENCE OF A MORDANT (2)

● Direct Staining ● Indirect Staining

● Gives color to the sections by using simple aqueous or alcoholic dye solutions.

Direct Staining

Example of Direct Staining (1)

Methylene Blue

● No mordant needed

Direct Staining

● Even without the use of a mordant, the stain can still color the tissue or cells.

Direct Staining

● Action of dye is intensified by using a MORDANT.

Indirect Staining

● Requires mordant

Indirect Staining

Example of Indirect Staining (1)

Hematoxylin

ACCORDING TO THE PRESENCE OF A DIFFERENTIATOR (DECOLORIZER) (3)

● Progressive Staining ● Regressive Staining ● Staining Method

● One particular example of staining method that utilizes progressive staining technique is H&E ○ H = hematoxylin ○ E = eosin

Progressive Staining

● Continuous application of dye ● Not differentiate and not decolorized the tissue

Progressive Staining

● When dye is taken up by the tissue, it is NOT decolorized.

Progressive Staining

● This H&E that applies progressive staining technique, this is the kind of use for frozen sections ○ No decolorization phase

Progressive Staining

● Requires a differentiator

Regressive Staining

● Tissue is first overstained, then afterwards the excess stain is removed or decolorized from unwanted parts of the tissue.

Regressive Staining

● Apply the stains first then use chemical that will decolorize certain parts that have been colored before

Regressive Staining

● Primary Stain = Basic ○ differentiator should be ________ solution

Acidic

Example: Regressive Staining

○ H&E ○ Acid alcohol

Differentiator for routine H&E

Acid alcohol

● Primary Stain = Acidic ○ differentiator should be an ________ solution

Alkaline

● Color of the dye is the color of the tissue after it is applied ○ Example: Red → Red

Orthochromatic Staining

● H&E Primary Stain = Hematoxylin (basic stain/dye) ○ differentiator should be _______

Acidic

● Whatever the color of the dye is, it is the expected color of the tissue.

Orthochromatic Staining

“Ortho” means “_______”

the same

● The color of the dye is different from the color of the tissue

Metachromatic Staining

(2) VITAL STAINING

● Intravital Staining ● Supravital Staining

Selective staining of living cell constituents

VITAL STAINING

● “Meta” means “_____” and also “________”

after and also change

● Color of the dye is not the color of the tissue after it was applied ○ Example: Red → Yellow

Metachromatic Staining

Intravital Staining (3)

○ Lithium ○ Carmine ○ India ink

● Injection of dye into any part of the animal’s body

Intravital Staining

Natural dye made up of alum and cochineal dye

Carmine

■ BEST vital dye (according to Gregorio’s)

Neutral red

● Stain is applied immediately after removal of tissue/organ from the body

Supravital Staining

● We removed the cells from the body and then outside that is where we applied the stain

Supravital Staining

Used in staining fungi

India ink

Used for the demonstration of mitochondria (mitochondria = organelle)

Janus Green B

Supravital Staining (5)

○ Neutral red ○ Janus Green B ○ Thionine (For frozen sections) ○ Toluidine blue ○ Nile blue

STAINING Flashcards

(85 cards)