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With the increasing demand for natural colorants as an alternative to synthetic stains, phytochemicals are gaining more attention. Natural stains from plant sources are cost effective, eco-friendly and bio-degradable. This study reveals the staining potential of red tamarind pulp extract (RTPE) on monocot and dicot stem tissue sections using bright-field and fluorescence microscopy. Phytochemicals were extracted from pre-dried immature red tamarind fruits using water and alcohol. Anthocyanins are characteristic red pigments and were observed more in both ethanol and methanol in comparison to its corresponding aqueous extract in water. Three excitation filters (Ultraviolet 330-380 nm; blue 450-490 nm; green 510-560 nm) and three mordants: potassium dichromate (K2Cr2O7), copper sulphate (CuSO4) and ferrous sulphate (FeSO4) were used to examine the plant tissue sections. RTPE with mordant elicits bright blue and orange to yellow fluorescence excitation of vascular bundles in violet and blue filter respectively. Hence RTPE can be deployed as a fluorescent dye to comprehend the plant cell component with higher precision and resolution.
Kraus JE, Arduin M. Manual básico de métodos em morfologia vegetal; 1997.
Ihuma J, Asenge G, Abioye J, Dick S. Application of methanolic extracts from Hibiscus sabdariffa Linn as a biological staining agent for some fungal species. IIJPAES. 2012;2(2):254-9.
Aguoru C, Okelionwu S, Olasan J. Value of Hibiscus rosasinensis petal extract as biological stain in plant systematic research. Journal of Chemical, Biological and Physical Sciences (JCBPS). 2016;6(2): 388.
Braide W, Akobundu C, Nwaoguikpe R, Njiribaeko L. The use of extracts from four local Nigerian plants for the staining of selected bacteria and moulds. African Journal of Microbiology Research. 2010; 5(1):79-86.
Suryawanshi H, Naik R, Kumar P, Gupta R. Curcuma longa extract–Haldi: A safe, eco-friendly natural cytoplasmic stain. Journal of Oral and Maxillofacial Pathology: JOMFP. 2017;21(3):340.
Horobin R, Kiernan J. Conn's biological stains: a handbook of dyes, stains and fluorochromes for use in biology and medicine: Taylor & Francis; 2020.
Bayazit ŞS. Investigation of Safranin O adsorption on superparamagnetic iron oxide nanoparticles (SPION) and multi-wall carbon nanotube/SPION composites. Desalination and Water Treatment. 2014; 52(37-39):6966-75.
Bordoloi B, Jaiswal R, Siddiqui S, Tandon A. Health hazards of special stains. Saudi Journal of Pathology and Microbiology. 2017;2(5):175-8.
Patil M, Shinde J, Jadhav A, Deshpande S. Adsorption of Methylene Blue in waste water by low cost adsorbent Rice husk. Adsorption. 2017;99:100.
Vutskits L, Briner A, Klauser P, Gascon E, Dayer AG, Kiss JZ, et al. Adverse effects of methylene blue on the central nervous system. The Journal of the American Society of Anesthesiologists. 2008;108(4): 684-92.
Bhuyan R, Saikia C. Extraction of natural colourants from roots of Morinda angustifolia Roxb.—Their identification and studies of dyeing characteristics on wool. Indian Journal of Chemical Technology. 2003;10(2): 131-136.
Deepali K, Lalita S, Deepika M, Stem M, Hibiscus FO. Application of aqueous plant extracts as Biological stains. International Journal of Engineering Science and Research. 2014;5(2):1586-1589.
Kumar N, Mehul J, Das B, Solanki J. Staining of Platyhelminthes by herbal dyes: An eco-friendly technique for the taxonomist. Veterinary World. 2015;8(11): 1321.
Siva R. Status of natural dyes and dye-yielding plants in India. Current Science. 2007;916-25.
Egbujo E, Adisa O, Yahaya A. A Study of the Staining Effect of Roselle (Hibiscus sabdariffa) on the Histologic Section of the Testis; 2008.
Abubakar S, Usman A, Etim V, Nnadi O, Alaku C. Application of organic dyes from roselle calyx (Hibiscus sabdariffa Linn) for mycological staining. Indian Journal of Innovations and Development. 2012;1(9): 687-90.
Raheem EM, Ibnouf A-AO, Shingeray OH, Farah H. Using of Hibiscus sabdariffa extract as a natural histological stain of the skin. American Journal of Research Communication. 2015;3(5): 211-6.
Verenkar NG, Sellappan K. Some potential natural dye yielding plants from the State of Goa, India. Indian Journal of Natural Products and Resources (IJNPR)[Formerly Natural Product Radiance (NPR)]. 2018; 8(4):306-15.
Shet Verenkar NG, Sellappan K. Evaluation of natural dyes Curcuma longa and Nyctanthes arbor‐tristis with different mordants on plant tissues under fluorescence microscopy. Microscopy Research and Technique. 2021;84(5):902-11.
Leong C-R, Azizi MAK, Taher MA, Wahidin S, Lee K-C, Tan W-N, et al. Anthocyanins from Clitoria ternatea attenuate food-borne Penicillium expansum and its potential application as food biopreservative. Natural Product Sciences. 2017;23(2):125-31.
Avwioro O, Aloamaka P, Ojianya N, Oduola T, Ekpo E. Extracts of Pterocarpus osun as a histological stain for collagen fibres. African Journal of Biotechnology. 2005;4(5):460-2.
Lakowicz JR. Principles of fluorescence spectroscopy: Springer Science & Business Media; 2013.
Albinsson B, Li S, Lundquist K, Stomberg R. The origin of lignin fluorescence. Journal of Molecular Structure. 1999; 508(1-3):19-27.
Donaldson L. Analysis of fibres using microscopy. Handbook of textile fibre structure: Elsevier. 2009;121-53.
Triol CB, Dionela ATV, Ecube Aid, Joy C, Mediodia A. The use of Clitoria ternatea (blue ternate) ethanolic extract as a potential stain for bacteria. Publiscience, 2020;3(1):42-47.
Vankar PS. Chemistry of natural dyes. Resonance. 2000;5(10):73-80.
Nazeri MA, Zain NM. Effect of different operating parameters on extraction of active compounds from pitaya peel by microwave assisted extraction (MAE). Jurnal Teknologi. 2018;80(2). Available:https://doi.org/10.11113/jt.v80.10974.
Shipp J, Abdel-Aal E-SM. Food applications and physiological effects of anthocyanins as functional food ingredients. The Open Food Science Journal. 2010;4(1):7-22.
Mayavel A, Nagarajan B, Muthuraj K, Nicodemus A, Prabhu R. Correlation and path coefficient analysis of selected red tamarind (Tamarindus indica var rhodocarpha) genetic resources. International Journal of Current Microbiology and Applied Sciences. 2018; 7(04):794-802.
Harborne J. Phytochemical methods: Methods of extraction and isolation. Chapman and Hall, London; 1998.
Kharbude B, Agarwal A. Different colour stains of different plant extract. Journal of Chromatography. 2000;347:447-52.
Rampriya S, Kumar NS. Characterization of Anthocyanins from Red Tamarind, Tamarindus indica var. rhodocarpa using Spectral Analysis. Indian Journal of Pharmacy and Biological Sciences. 2019; 9(3):926-931.
Giusti MM, Wrolstad RE. Characterization and measurement of anthocyanins by UV‐visible spectroscopy. Current Protocols in Food Analytical Chemistry. 2001(1):F1. 2.1-F. 2.13.
Priyadarsini KI. The chemistry of curcumin: from extraction to therapeutic agent. Molecules. 2014;19(12):20091-112.
Chukwu O, Odu C, Chukwu D, Hafiz N, Chidozie V, Onyimba I. Application of extracts of Henna (Lawsonia inamis) leaves as a counter stain. African Journal of Microbiology Research. 2011;5(21):3351–3356.
Bassey RB, Oremosu AA, Osinubi A. Curcuma longa: Staining effect on histomorphology of the testes. Macedonian Journal of Medical Science. 2012;5(1): 26-9.
Saiki P, Thitipramote N. Extraction of natural histological dye from black plum fruit (Syzygium cumini). Journal of the Microscopy Society of Thailand. 2011;4(1): 13-5.
Adeyemo FE, Singh G, Reddy P, Stenström TA. Methods for the detection of Cryptosporidium and Giardia: from microscopy to nucleic acid based tools in clinical and environmental regimes. Acta Tropica. 2018;184:15-28.
Kubista M, Sjöback R, Eriksson S, Albinsson B. Experimental correction for the inner-filter effect in fluorescence spectra. Analyst. 1994;119(3): 417-9.