Hendirikka (Mirabilis jalapa) Derived Dye: An Alternative Natural Stain for Human Red Blood Cells.

Abstract

Background: Staining is a vital technique in clinical microscopy that enhances the contrast of cellular elements during morphological examinations. Although synthetic dyes are extensively used in staining biological samples, they were reported to be associated with potential adverse effects on laboratory personnel and the environment. Recent studies have extensively documented the possible usage of plant-derived dyes in staining biological tissues due to their affordability, safety and biodegradability. Objective: To evaluate the staining potential of a natural dye from Mirabilis jalapa on human red blood cells (RBCs) Methods: This was a laboratory-based experimental study. The magenta colour flowers of M. jalapa were collected and cleaned with tap water. Then, they were subjected to drying in an oven at 40°C and ground into a dry powder. 20 g of dry plant material was mixed with 100 mL of double distilled water and incubated at 70°C in a water bath for 4 hours and subsequently at 25°C for 12 hours. The extract was filtered through gauze and centrifuged at 5000 rpm for 15 min. The supernatant was filtered using Whatman No.1 filter paper, evaporated by a rotary evaporator and redissolved in distilled water. A serial concentration of dye solution was prepared and applied for 10 min on blood smears fixed by absolute methanol and observed under oil immersion. The photographs taken by a digital microscope were subjected to intensity measurement using ImageJ version 1.54f and statistically compared with Leishman stain by unpaired t-test using SPSS version 18.0. Results: Results demonstrated that dyes extracted from M. jalapa exhibit a staining potential on human RBCs in a concentration-dependent manner. Furthermore, the ImageJ analysis revealed that 80 mg/mL of M. jalapa dye has the potential to give optimal staining on RBCs with an intensity of 116.82±0.53, which is almost comparable to Leishman stain (I=116.43±0.29, p=0.151). Conclusion: The dye obtained from M. jalapa demonstrated a staining potential on human RBCs; however, further purification studies are recommended to establish this dye as an alternative natural stain for RBCs.