Kleinenberg’s hæmatoxyline.—This formula differs from the previous one in being an alcoholic solution. The calcium chloride is added because it “sets up diffusion currents between the alcohol in the material to be stained and the alcoholic staining solution, so enabling the latter to penetrate more rapidly” (Squire). It is much used in staining embryonic specimens in bulk before embedding in paraffin, and was strongly recommended for that purpose by Foster and Maitland Balfour.
Various formulæ have been given from time to time. That advised by Squire (Methods and Formulæ, p. 25) can be accurately made up without much difficulty.
| (a) | Crystallised calcium chloride | 20 | grs. | 1/2 | oz. |
| Distilled water | 10 | c.c. | 2 | drms. | |
| (b) | Alum | 3 | grms. | 32 | grs. |
| Distilled water | 16 | c.c. | 170 | mins. |
Mix and add
| Rectified spirit | 240 | c.c. | 8 | ozs. |
Allow it to stand and any excess of calcium sulphate, &c., to separate. Filter and add
| Hæmatoxyline | 2 1/2 | grms. | 25 | grs. |
A little thymol should be added as a preservative.
In making up these solutions care must be taken that only distilled water is used, and that all the vessels employed have been previously rinsed out with it, otherwise precipitation of the hæmatoxyline will occur.
Should sections be overstained in hæmatoxyline, this may be remedied by washing it in a half per cent. solution of acetic acid, until sufficient of the stain is discharged, but the staining is more diffuse than if the happy mean had been hit in the first instance.