The Determination of Optimum Staining Time for Hibiscus Extract Nuclear Staining Using Buffered Formalin Fixed, Paraffin Embedded Lung Tissue

Abstract

This exploratory study investigated the optimum nuclear staining time of H. sabadariffa aqueous extract mordanted with ferric chloride solution on 10% neutral buffered formalin fixed, paraffin embedded lung tissue. Blocks of lung tissues were retrieved from the post-mortem block archive of the Pathology Department, University of Ilorin Teaching Hospital, Ilorin, Kwara State, Nigeria. The blocks were sectioned at 3-5 microns with a Leica Rotary microtome and stained with Hibiscus-Eosin staining technique for 5 minutes (A), 10 minutes (B), 15 minutes (C), 20 minutes (D), 25 minutes (E), 30 minutes (F) and 60 minutes (G). Parallel staining was done with H&E as control. Stained slides were blindly assessed independently by three Histopathology experts for cell membrane appearance, nuclear membranes, cytoplasm transparency and extracellular matrix. Photomicrographs revealed satisfactory staining by the Hibiscus-eosin technique comparable to standard H&E.  Assessment by the Histopathology experts revealed that optimal staining was achieved between 5 -10 minutes. The optimum staining time for Hibiscus nuclear staining is 5-10 minutes.