This overview of slide staining originally appeared on wardsci.com.
The exact chemistry involved in most staining procedures is unknown. There is probably more than one type of chemical interaction occurring during any staining procedure. Interactions can be electrostatic, covalent, or based on physical properties that affect stain penetration, including texture, density and solubility.
One prevalent mechanism is thought to be the electrostatic interaction of the chromogen with tissue molecules leading to the classification of many stains as acidophilic or basophilic. Basic dyes color acidic molecules - for example, hematoxylin, stains DNA (an acid) dark blue. Acid dyes color more basic molecules - for example, eosin, makes most cytoplasms red/pink.
Choose among the available stains to highlight structures of interest.
| What are you looking for/at? | Commonly used stain | Examples of what you see: |
| Bacteria | Gram’s Iodine Gram’s Stain Kit |
Violet - positive Red - negative |
| General plant tissue | Quadruple stainSafranin OFast GreenCrystal Violet
Orange G |
Red nuclei, chromosomes, lignified cell wallsGreen cellulose cell wallsPurple starch grainsYellow acidophillic cytoplasm |
| General animal tissue | H&E(Hematoxylin & Eosin) | Blue nuclei (H) Pink cytoplasm and connective tissue (E) |
| Dramatic cell and tissue features | Mallory triple | Blue cartilage and collagen Red nucleus Orange cytoplasmic elements |
| Dramatic cell and tissue features | Masson | Green collagen and mucus Red nucleus Brown-Black chromatin |
| Standard Blood smears | Giemsa or Wright’s | Orange/Red erythrocytes Blue/Purple lymphocytes Multi-color granulocytes |
| Mitotic figures | Iron Hematoxylin | Blue/Black chromosomes, cytoskeleton (tubulin), and mitochondria |
| Fats and lipids | Osmium tetroxide or Sudan Black | Black lipid droplets, cell membranes, myelin |
| Mucus/complex polysaccharides | PAS (Periodic Acid/Schiff) |
Fuschia/Red secretory granules in goblet cells |
| Neurofibrils | Silver (multiple methods) | Black neuronal cytoskeleton |
| Elastic connective tissue | Orcein or Verhoeff | Brown/Black elastin containing fibers |
Also see: Basic Stain Set and Vital Stain Set
WARD’S complete stain guide is also available in PDF format.
Related Articles: Also see our Slide Preparations Guide.
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