| Azure II-eosin | 3·0 | grm. |
| Azure II | 0·8 | " |
| Glycerine, pure | 125·0 | " |
| Methyl alcohol, pure | 375·0 | " |
The film is first fixed in absolute alcohol. The proportion of stain usually used is one drop of stain to 1 c.c. of water. Stain for about ten minutes and then wash in water.
The details of the application of the Giemsa stain to films fixed wet and to sections must be sought in larger works on technique. These works should also be consulted for information regarding the use of Pappenheim’s Panchrome mixture.
There are numerous formulae of stains containing ripened Hæmatoxylin or its essential principle, Hæmatein. A mordant is necessary, one of the alums being usually employed. The mordant may be included as an ingredient in the staining mixture, or it may be used separately as in the case of the so-called iron-hæmatoxylins, wherein ferric ammonium alum is used separately and is followed by staining with hæmatoxylin or hæmatein. A few of these stains of general application may now be mentioned.
Delafield’s (or Grenachier’s) Hæmatoxylin.
| Hæmatoxylin crystals | 4 | grm. |
| Absolute alcohol | 25 | c.c. |
| Saturated aqueous solution of ammonia-alum | 400 | " |
Mix these ingredients, and leave exposed to light and air for three to four days. Filter and add—
| Glycerine | 100 | c.c. |
| Methyl alcohol | 100 | " |
Allow the mixture to stand until the colour is sufficiently deep, then filter and place in a stoppered bottle. The solution should be allowed to ripen for at least two months before use. Dilute aqueous solutions of the stain are of service for films and for sections. A trace of acetic acid may be added at the moment of use, for sharp differentiation.
Ehrlich’s acid hæmatoxylin, Mayer’s hæmalum, and Mayer’s glychæmalum are also useful. Their formulæ will be found in larger works.