Histomorphological Staining of Selected Organs by Iron-Roselle Nuclear Technique and Natural Counter Stains

Background: Roselle extract is proving to be a staining alternative to haematoxylin in histopathological studies. Early studies concentrated on organs like lymph nodes, appendix, liver, kidneys and brain. The present study aims at expanding the frontiers of knowledge on the histological application of natural dyes from H. sabdariffa on the histo-morphology of some connective tissue rich organs viz uterus, ovary, breast and skin. Electrostatic compatibility with natural counter stains such as Sorghum bicolour, Baphia nitida (Camwood) and comparative reactions with standard counter stains like eosin, Masson Trichrome and Van Gieson was also explored.

Methods: Dry roselle calyces, Sorghum bicolour stalk, Camwood (Baphia nitida) powder were purchased at local markets in Ilorin, capital of Kwara State, Nigeria and identified at the Department of Plant Biology, University of Ilorin, Kwara State, Nigeria. Staining solutions were prepared as recommended. Blocks of tissues from the Skin, Ovary, Breast and Uterus were retrieved from the archive of the Pathology Department, University of Ilorin Teaching Hospital, Ilorin. Six sections were cut at 4 microns from each organ for each of the following Hibiscus based techniques-{Hib-Eosin (A), Hib-Trichrome(B), Hib-Van Gieson(C), Hib-Sorg (D), Hib-Camwood(E)} and haematoxylin based staining techniques { H&E(F), MT(G), HVG(H), Hx-Sorg(L) and Hx-Camwood (J)}as parallel controls. Stained sections were blindly evaluated by three independent Histopathology experts for nucleus-cytoplasm staining, connective tissue demonstration and morphological preservation. Statistical evaluation was done using Near test, Kruskal-Wallis test and Kappa statistics with p-value set at 0.05.

Results: Histological observations show a satisfactory morphological demonstration of all the organs. Statistical analysis shows no significance difference between the Hibiscus sabdariffa based staining techniques and standard haematoxylin based staining techniques.

Conclusion: The histo-morphological demonstration of skin, ovary, uterus and breast was well preserved by Hibiscus sabdarifa solution (Iron-Roselle) nuclear staining and showed satisfactory electrostatic combinations with eosin, trichrome, Sorghum bicolour, Van Gieson and Camwood counter stains.