Staining Flashcards
Staining
Process of coloring the different structural components to be more visible through variation of colors promoting easier differentiation and identification.
True or False. Dye is a benzene ring composed of Chromophore and Auxochrome. Chromophore gives the dye affinity to attach on tissues while Auxochrome is the Coloring component.
1st statement: True
2nd statement: False
Chromophore - coloring Component
Auxochrome - affinity to attach tissues.
Active coloring agent of Hematoxylin
Hematein
Hemaitin
Formed by the oxidation of Hematoxylin
Most frequently used in combination with Hematoxylin
ALUM
IRON
CHROMIUM
COPPER SALTS
Process which the action the of the dye is intensified by adding another agent or a MORDANT
Indirect Staining
H & E Staining Method is what type of staining process
Indirect, Regressive Staining Process
Routinely used COUNTERSTAIN after Hematoxylin and before MB
EOSIN
Give the Color of the structure.
- Nucleus
- Cytoplasm
Nucleus. Hematoxylin stains nucleus to BLUE, therefore basophilic. Since nuclear structures are acidic, negatively charges therefore, high affinity to basic stain.
Cytoplasm. Eosin Y stains cytoplasm to RED/PINK, “acidophilic”. Cytoplasmic structures are basic and with high affinity to acidic dye, therefore retaining the color of Eosin.
Purpose of Acid alcohol
Differentiating agent. Rinses or remove stains in cytoplasm while nucleus remains stained by giving elaborate and sharp detail of the nucleus. It also removes hematoxylin not bounded to the mordant.
Purpose of Ammonia Water
Bluing agent. Converts soluble red color of the nucleus by hematoxylin to an insoluble blue color.
Enumerate the arranged steps in H & E staining
- Xylol (I/II/II) – deparaffinization
- Descending grade of alcohol (95>90>80>70)
- Water
- Stain: Hematoxylin
- Water
- Acid alcohol
- Water
- Ammonia water
- Wash
- Stain: Eosin Y
- Ascending grade of Alcohol (70>80>90>95)
- Xylene (I/II)
- Mount and Label
Give their functions and purpose:
- Xylol (I/II/II) – deparaffinization
- Descending grade of alcohol (95>90>80>70)
- Water
- Stain: Hematoxylin
- Water
- Acid alcohol
- Water
- Ammonia water
- Wash
- Stain: Eosin Y
- Ascending grade of Alcohol (70>80>90>95)
- Xylene (I/II)
- Mount and Label
- Xylol (I/II/II) – deparaffinization; remove excess paraffin wax
- Descending grade of alcohol (95>90>80>70): Rehydration; Prevent damage and detachment of tissues
- Water: Remove excess agents
- Stain: Hematoxylin. Stains Nuclear Structure
- Water: Decolorizer
- Acid alcohol. Differentiating agents
- Water: Remove excess alcohol
- Ammonia water. Bluing Agent
- Wash
- Stain: Eosin Y. Stain Cytoplasmic component
- Ascending grade of Alcohol (70>80>90>95): Dehydration to maintain cell structure and shape.
- Xylene (I/II): Clearing excess alcohol
- Mount and Label
Precautions in Staining
- Stains in skin should be avoided
- Use new/fresh albumin adhesive to prevent
detachment of sections from the slide - Check reagents
- Prevent blurred sections under microscope
Identify the colors demonstrated.
Nucleus Cytoplasm Karyosome RBCs, EOS, Keratin Plasma Cells, Osteoblast Cartilage Calcium and Calcified Bones Decalcified Bone Muscle Fibers, Thyroid Colloid and Elastic Fibers
Nucleus: BLUE/BLACK Cytoplasm: PINK Karyosome: DARK BLUE RBCs, EOS, Keratin: BRIGHT RED-ORANGE Plasma Cells, Osteoblast: PURPLE Cartilage: PINK/LIGHT BLUE/DARK BLUE Calcium and Calcified Bones: PURPLISH BLUE Decalcified Bone: DEEP PINK Muscle Fibers, Thyroid Colloid and Elastic Fibers: DEEP PINK
