Heidenhain's Iron Hematoxylin Variants

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.

Hover the cursor over a contraction for more information. 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



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