Table 2.
Results of 25 histochemical stains, modified from the cited references, applied to bone tissue from porcine models of haematogenous and implant associated osteomyelitis induced with Staphylococcus aureus S45F9.
| Stain | Staining pattern | Bacteria color | Matrix | Contrast | Protocol Reference |
|---|---|---|---|---|---|
| Connective tissue stains | |||||
| Alkaline congo red | Red: Amyloid (via. H-bonds), elastic fibers, eosinophil granules Blue: Nuclei |
Purple | No | ++ | 15 |
| Crystal violet | Red-purple: Amyloid, mucin, renal hyaline Blue: Background |
Purple | Yes Purple |
++ | 15 |
| Gomori's reticular fibers | Black: Reticular fibers Gray: Nuclei |
Brown/purple | No | + | 15 |
| Luna | Red: Eosinophils, RBC (red blood cells) Blue: Background |
Blue | Yes Blue |
+++ | 16 |
| Martius scarlet blue | Blue: Nuclei, collagen, (old fibrin) Yellow: RBC, (early fibrin) Red: Muscle, fibrin |
Purple | No | + | 15 |
| Masson-trichome | Blue/black: Nuclei Red: Cytoplasm, muscle, RBC Blue: Collagen |
Brown/purple | No | ++ | 15 |
| Picro-sirius red | Red: Collagen type 1-3, keratohyalin granules Polarized light: Collagen 1 - yellow/orange/red, collagen 3 - green |
Orange | Yes Orange |
++ | 17 |
| PTAH | Dark blue: Muscle striations, neuroglia, fibrin, amoebae Blue: Nuclei, cilia, RBC Light blue: Myelin Deep red-brown: Collagen, osteoid, cartilage, elastic fibers Pale pink-brown: Cytoplasm |
Brown/purple | No | + | 16 |
| Van Gieson | Blue/black: Nuclei Red: Collagen |
Brown | No | + | 15 |
| Verhöeff's | Black: Elastic tissue fibers | Purple | No | + | 15 |
| Carbohydrate stains | |||||
| Alcian blue ph1 | Blue: Acid mucins, proteoglycans, hyaluronic acid Red: Nuclei |
Blue | Yes Blue |
+ | 15 |
| Alcian blue ph3 | Blue: Acid mucins, proteoglycans, hyaluronic acid Red: Nuclei |
Blue | Yes Blue |
+++ | 15 |
| PAS | Magenta: Glycogen, neutral/sialomucins, glycoproteins Blue: Nuclei |
Purple | Yes Purple |
++ | 15 |
| Safranin O | Black: Nuclei Grey/green: Cytoplasm Orange/red: Cartilage, mast cells |
Purple | Yes Purple |
+++ | 18 |
| Toluidine blue-acetone | Blue/purple: Mast cell granula Blue: Nuclei and background |
Blue | Yes Blue |
++ | 15 |
| Lipid stains | |||||
| Oil-red O | Red: Fat Blue: Nuclei |
Purple | Yes Purple |
++ | 15 |
| Pigment and mineral stains | |||||
| Perls' Prussian blue | Blue: Ferric iron Red: Nuclei |
Red | No | ++ | 15 |
| Von kossa | Black: Mineralized bone Red: Osteoid |
Brown | No | + | 15 |
| Alizarin red S | Orange-red: Calcium deposits (especially small deposits) | Yellow/brown | No | +/++ | 15 |
| Microorganism stains | |||||
| Giemsa | Dark blue: Protozoa, microorganisms Pink-pale blue: Background Blue: Nuclei |
Blue | Yes Blue |
+++ | 15 |
| Grocott methenamine-silver | Black: Fungi, pneumocystis, melanin, hyphae, yeast form cells of fungi Taupe-dark gray: Mucins, glycogen Pale green: Background |
Black | No | ++ | 15 |
| Levaditis | Black: Spirochaetes, some organisms and fungi Yellow: Background |
Black | No | ++ | 16 |
| Gram | Blue/black: Gram positive bacteria Red: Gram negative bacteria |
Purple/blue | Yes Purple/blue |
+++ | 15 |
| Nucleic acid stains | |||||
| Feulgen nuclear reaction for DNA | Red/purple: DNA Green: Cytoplasm |
Turquoise | No | + | 15 |
| Methyl green-pyronin | Green/blue: DNA Red: RNA |
Red/pink | Yes Red/pink |
+++ | 15 |
Contrast: poor contrast (+), fair contrast (++), good contrast (+++) between bacteria and background.