Staining Flashcards by John Lesther Alonsagay

Example of direct staining

Methylene blue, Eosin

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In direct staining, sections are stained with simple __ or __

Aqueous, alcoholic solutions

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Indirect staining: M and A

Mordant and Accentuator

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The bridge for IS

Mordant

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Accelerator of staining for SI

Accentuator

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Ex. of mordant

K alum (Ehrlich) and Iron (Weiger’s)

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Ex. of Accentuator

K hydroxide, phenol

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Follows a definite sequence

Progressive staining

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Does not wash or decolorize

Progressive staining

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Differentiation relies on the selective affinity of dyes

Progressive staining

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Les favored than regressive

Progressive staining

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Tissue is first overstained; decolorize

Regressive staining

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Differentiation/ decolorization selective removal of excess stain

Regressive staining

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Ex. of Regressive staining

Gram stain Acid stain

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Forms a soluble salt w/ the metal so that the latter is dissolved out

Acid differentiator

ex. HCI; HaC

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Oxidizes the dye to a colorless substance

Oxidizing Differentiator

Ex. K ferricyanide / K Permanganate

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If the primary stain is basic, the decolorizer is aciding, and vice-versa

Mordant Differentiator

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___ acts a differentiator for both basic and acidic dye

Alcohol

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Differentiation is also done to restain faded slide

Mordant Differentiator

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Stains tissue with acolor that is different from that of the stain itself - Metachromasia

Metachromatic staining

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Metachromatic staining is for

cartilage, connective tissue, epithelial mucins, mast cell granuels and Amyloid

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Ex of Metachromatic staining

Toluidine blue - red purple on mast cell

Cresyl blue - blue green on reticulocytes

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Application of a different color or stain to provide contrast and background staining

Counterstaining

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Counterstaining ex.

Cytoplasmic and Nuclear

Cytoplasmic: eosin Y, Picric acid, Lissamine Green

Nuclear: Neutral red, hematoxylin, Safranin O

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General relationship of tissues and cells

Microanatomical staining

Structures found in the cytoplasm and nucleus

Cytoplasmic staining

Bacterial morphology

Negative staining

Demonstration of tissue elements by the use of metallic salts (colorless solutions)

Metallic impregnation

Not absorbed by tissue just like stain; rather held physically on the surfaces as a precipitate as a reduction product in certain tissue components

Metallic Impregnation

Selective staining of living cells

Vital staining

Rough Endoplasmic Reticulum

Trypan blue

Mitochondria

Janus Green

Nucleus is not demonstrated

Vital staining

Done by injecting the dye into any part of animal body (intravenous intraperitoneal, subcutanous)

Intervital

Ex of intervital

Lithium, carmin and india ink

Stains living cells immediately after removing from the living body

Supravital

Ex of Supravital

Neutral red, Janus green, trypan blue

Two categories of dyes

Natural dyes and Synthetic Dyes

Ex of Natural Dyes

Hematoxylin, Cocchineal dyes, orcein, Saffron

Ex of synthetic dyes

Aniline or Coal Tar dyes

Hematoxylin is derived from

Hematoxylin campechianium

Most valuable stain used

Hematoxylin

Active coloring agent in hematoxylin, Exposure to: __ or __

Hematin; Air/sunlight or oxidizing agents

Alum Hematoylin

Progressive staining

Alum hematoxylin is counterstained with?

Eosin, Congo Red, Safranin

Iron Heamtoxylin

Differentail of regressive staining

Spermatogenisis

Copper hematoxylin

For fresh material and smear prep

Cochineal Dyes

Cochineal dyes are derived from

female coccus cacti

Cochineal dyes are treated w/ __:__

Alum: Carmine

Neuropathological Studies

CD w/ Picric Acid

Glycogen Demonstration

CD w/ Aluminum Chloride

For elastic fiber

Orcein

Orcein is derived from

Lichens; vegetable dye

Orcein is treated with?

Ammonia and exposed to air (blue/violet color)

Lime and soda

Litmus

Used for indicator

Orcein

Coal tar dyes is derived from

Hydrocarbon benzene

dye appears in the tissue

Chromophores

Gives color to the dye

Chromogens

Retains color; permanent

Auxochrome

what is the base of acid dye

sodium

base of basic dye?

HCl

Neutral dye is __

Soluble in alcohol but not in water

For regressive staiining

Ehrlich's Hematoxylin

Cartilage and cement lines of bones: BLUE

Ehrlich's hematoxylin

Staining time of Ehrlich's Hematoxylin

15-40 mins

Routine nuclear staining

Harris Hematoxylin

Exfoliative cytology

Harris Hematoxylin

Sex Chromosomes

Harris Hematoxylin

Staining time or Harris Hematoxylin

5-20 minutes

Routine pruposes

Coles hematoxylin

Used in sequence w/ Celestine blue

Coles hematoxylin

Coles hematoxylin staining time

10 mins

Mucopolysaccharide

Mayer's Hematoxylin

Nuclear Staining

Mayer's Hematoxylin

Demonstrating muscle fibers and connective tissues

Weigert's Hematoxylin Solutions

Recommended when the preceding stains contain acid

Weigert's Hematoxylin Solutions

Cytological stain recommended for regeressive staining of thin sections

Heidenhain's Hematoxylin

Demonstration of chromatin, Chromosomes, nucleoli, centrosomes, and mitochondria

Heidenhain's Hematoxylin

Voluntary muscle striations and myelin

Heidenhain's Hematoxylin

Demonstrates structures in paraffin, celloidin and frozen section

Phophotungstic acid hematoxylin (PTAH)

in Phophotungstic acid hematoxylin (PTAH), Collagen, bone, cartilage is colored ___

Orange-red or Brownish red to deep brick red stain

in Phophotungstic acid hematoxylin (PTAH), Nuclei, fibrin, muscle striations, myofobrils, fibroglia is colored __

Blue

Phophotungstic acid hematoxylin (PTAH) staiing time __

12-24 hrs

Staining connective tissue and cytoplasm differentially

Eosin

Counterstaing after hematoxylin and before methylene blue

Eosin

Background for constrasting stains

Eosin

2 Shades of Eosin

1. Bluish (Eosin B) - deeper red color 2. Yellowish (Eosin Y) - Green yellow flourescence

Basic nuclear stain; diagnosis of Diptheria; Fresh sputum; Aerogenes bac

Methylene blue

MB is heated with fixed alkali/ alkali carbonate

Methyl Violet

Sub for Thionine if FFT

Toluidine blue

for nissle granules

toluidine blue

Amyloid in FFT; platelets

Crystal violet

Counterstaining of epithelial sections

Aniline blue

Plasma stain for acid-fast organism

Basic Fuchsin

Ascaris eggs and erethrocyte; bacterial spore stain.

Malachite green

both decolorizer and counterstain

malachite green

Contrast stain for Gran's staining

Bismarck brown

Utilized for manufacture of paints

Prussian blue

Acid mucopolysaccharide

Alcian blue

Embryos

Congo red

mitochondria

Janus Green

Substitute for carbol fuchsin

Night blue

Phospholipids, neutral lipids ___% solution boiled in ___% what?

Sudan Black; 0.5%; 70%

Neutral lipids only ___% ___ in ___% Alcohol

Sudan IV 2% benzoic acid in 70% alcohol

Fat stain for CNS

Sudan III

Water should always be?

Distilled

alcohol used are ___ & ___

Ethyl Alcohol and Methyl Alcohol

Aniline Water -- __cc added to every ___ to 1 liter of hot DW

10cc; 1/2

Phenol - aqueous solution of ___-___%

0.5 - 5 %

Holds 5-9 slides

Coplin Jar

Holds 5-19 Slides

Slotted staining dishes

Holds 10-30 slides

Metal/glass staining racks/ carriers

Char of a good mounting medium

- Refractive index near of the glass (1.518) - Should not dry quickly - Should not dissolve out tissue sections - Should not cause shrinkage and distortion -Should be permanent

- water miscible preparations

Aqueous Media

RI of glycerin gelly

RI: 1.47

RI of Gum Arabic (Farrant's medium)

RI: 1.43

Apathy's medium

RI: 1.52

Preparations that have been dehydrated and cleared in Xylene and Toluene

Resinous Media

Natural and synthetic resins

Resinous media

Canada Balsam RI

1.524

DPX RI

1.532

Xam RI

1.520

Permount RI

1.518

Clarite RI

1.544

Process of sealing the margins of the coverslip to prevent escapage of fluid

Ringing

Ex of ringing

Kronig Cement

Kronig Cement: __ parts of paraffin wax mixed with ___ parts ___

2 parts of PW mixed w/ 4-9 parts POWDERED COLOPHONIUM RESIN (heated and filtered)