Iron Hematoxylin
Variants of Heidenhain's method
Heidenhain's iron hematoxylin technique uses two seperate solutions for staining. First an iron mordant is applied, usually ferric ammonium sulphate or ferric chloride. This is followed by a hematoxylin solution. The overstained preparations are then differentiated in the iron mordant solution, often diluted, or in some similar solution. Numerous variations on this theme have been published, most of them staining very similarly, as they differ largely in the concentration of the reagents and the times for which they are applied.
In the chart below it is to be understood that all solids are in grams and all fluids in millilitres. The amounts given are to be dissolved in 100 millilitres of the solvent for each solution. The solvent is distilled water unless otherwise stated.
Placing the mouse cursor over any of the contractions in a solution column should cause the formula of the solution to be displayed.
The term "visual" in the "Diff" column means to control differentiation microscopically.
| Variant | Sol. A | Sol. B | Sol. C | Time A | Time B | Diff | Comment |
| Heidenhain | IA 2.5 | HX 0.5 | – | ½ - 24 hrs. | ½ - 24 hrs. | A, visual | |
| Bütschli | FA 2 | HX 0.5 | – | 24 hrs | 3 hrs | – | For protozoa |
| Diamond | IA 4 |
HX 0.5 Ter 0.1 |
PA sat. alc. | 5 min. | 5 min. | C, 3-5 min. | |
| Dobell | IA 1 70% Eth |
HN 1 70% Eth |
HCl 0.1 70% Eth |
10 min | 10 min | C, visual | For protozoa |
| Freitas | IA 0.5 70% Eth |
HN 1 70% Eth 60 PB 30 |
PA 0.3 70% Eth |
1 hr | 1 hr | C, visual | |
| French | IA 3.5 70% Eth |
HX 1 95% Eth 98 LC sat 2 |
IA 1 | Overnight | Overnight | C, visual | |
| Galiano | IA 3 |
HX 1 AC 20 |
EO 1.5 AC 25 |
15 mins | until dark | C, visual | Rinse in NH Eth |
| Haggquist | IA 5 | HX 1 | FC 1 | 1 hr | 1 hr | C, visual | |
| Hance | IA 2.5 | 10% HX 10 2.5% SB AD 100 |
½ - 2 hrs | 30 min | C, visual | Add SB until sol B changes from yellow to a plum colour |
