Outline

1. Adhesives
2. General principles of staining
3. H&E
4. Mounting
 Adhesives
1. Mayer’s egg albumin
2. Dried albumin
3. Gelatin
4. Starch paste
5. Plasma
6. Celloiden
7. Chrome alum adhesive
• Processes involved:
– Osmosis, solvation, adsorption,
absorption
• Chemical properties involved:
– Affinity (pH)
• Types of stains:
– Natural dyes
– Synthetic dyes
• chromophore!
• General
Methods/terms:
1. Direct 10. Histological
2. Indirect 11. Metallic
3. Progressive impregnation
4. Regressive 12. Vital staining
5. Metachromatic 13. Supravital
6. Counterstaining/ 14. Intravital
differentiation
7. Microanatomical
8. Histochemical
• Pre-staining considerations:
1. Precautions & proper technique
2. Type of tissue
3. Component of interest
4. Prior reagent used:
1. Deparaffinization (*sections to water)
2. Collodionization / parcelloiden / celloidin
3. Frozen sections
4. Decalcifying agents
 Restaining

Removal of coverslip:
Xylene – 30 min
Xylene – 24 hrs OR heating

Oxalic acid – 5 min

Repeat washing

KMnO4 – 5-10 min

 Mounting
• Characteristics of a good mountant?
• Ringing?
• Aqueous media
– Water (w/ glycerine & jelly)
– Glycerin
– Gum arabic (Farrant’s medium)
– Karo corn syrup
• Resinous media
– Canada balsam
– DPX
– Xam
– Permount
– Clarite
Mounting
 H&E
• Nuclei - blue - with some
metachromasia
• Cytoplasm - various shades of pink-
identifying different tissue
components