A study of various modifications of Lawsonia inermis (Henna) leaf extract as a cytoplasmic stain in liver biopsies

James Olayiwola Adisa, Khadijah Khalid Musa, Ejike Chukwudi Egbujo, Ijeoma Matinda Uwaeme


Background: The leaves of Lawsonia inermis (henna) is widely used as cosmetic agent for body art to colour the hair, skin and nails, and hence is considered as a candidate stain for tissues.  Lawsonia inermis leaves contain a pigment which is the source of red-brown dye. The application of natural dyes for staining biological tissues will reduce cost and is more environmentally friendly.

Methods: The natural dye Lawson (2-hydroxy-1,4-napthaquinone) was extracted from the dry leave of Henna using two solvents (distilled water and ethanol). The two extracts were formulated into various staining solutions at different concentrations. The staining solutions were modified with potassium alum and phenol. Liver sections were stained using haematoxylin and eosin method alongside various modifications of the plant extract as counterstain at different timing and temperature conditions.

Results: The solutions were found to be acidic in nature and stained the cytoplasm golden brown with a well-defined cytoplasmic boundary of hepatocytes. The best staining effect was obtained with 1% concentration at room temperature for 10 minutes. This provides a measure of contrast between the nuclei, cytoplasm and connective tissue that is adequate for diagnosis. Phenol used as an accentuator has been found to improve the staining but caused swelling of cell. Mordanting with potassium alum did not cause swelling but did not give the kind of resolution as obtained with Phenol. Use of the henna extract at 60oC gave the poorest result which showed that high temperature adversely affect the staining reaction by presenting almost the same color to the structures irrespective of the solvent or concentration.

Conclusions: This findings suggest that Henna leave extract can be used as a suitable cytoplasmic connective tissue stain.


Counterstain, Cytoplasmic stain, Lawsonia inermis, Natural dyes, Stain

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