Clitoria Ternatea) to Stain Blood Caused Scientists to Concern About Using the Cells in Different Animal Peripheral Blood Natural Dyes Instead of Synthetic Dyes

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Clitoria Ternatea) to Stain Blood Caused Scientists to Concern About Using the Cells in Different Animal Peripheral Blood Natural Dyes Instead of Synthetic Dyes บทความวิชาการ สัตวแพทยั มหานครสาร์์ JOURNAL OF MAHANAKORN VETERIINARY MEDIICIINE Available online: www.vet.mut.ac.th/journal_jmvm การใช้สสกี ัดจากพืชในการย้อมสีเนื้อเยื่อ 1,# 2 อารยา สืบขาเพชรํ และณฐกาญจนั ์ นายมอญ 1ภาควิชากายวิภาคศาสตร ์ คณะสัตวแพทยศาสตร์ มหาวิทยาลัยเทคโนโลยีมหานคร กรุงเทพฯ 10530 2ภาควิชาเทคนิคการสัตวแพทย์ คณะเทคนิคการสัตวแพทย์ มหาวิทยาลัยเกษตรศาสตร์ กรุงเทพฯ 10900 บทคัดย่อ: ปัจจุบันการศึกษาทางจุลกายวิภาคศาสตร์ได้มีการใช้สีย้อมมากมายหลายชนิดเพื่อย้อมเนื้อเยื่อต่างๆ ซึ่งขึ้นกับวัตถุประสงค์ในการศึกษา สีที่ใช้ย้อมเนื้อเยื่อโดยทั่วไปมีแหล่งที่มาทั้งจากแหล่งธรรมชาติและจากการ สังเคราะห์ ด้วยในปัจจุบันทั่วโลกให้ความสําคัญในการใช้สารต่างๆ ที่ไม่มีผลกระทบต่อสุขภาพของมนุษย์และ เป็นมิตรกับสิ่งแวดล้อม การใช้สีธรรมชาติจากพืชในการย้อมเนื้อเยื่อจึงได้รับความสนใจมากขึ้น เนื่องจากส่วน ต่างๆ ของพืชนั้นมีส่วนประกอบของสีอยู่มากมาย อีกทั้งยังมีราคาถูกกว่าและสามารถใช้งานได้อย่างยั่งยืนกว่า สีที่ได้จากการสังเคราะห์ รายงานฉบับนี้ได้ทวนสรุปโดยย่อเกี่ยวกับการแบ่งชนิดของสีธรรมชาติจากพืชในแง่ ของโครงสร้างทางเคมีของสี วิธีการสกัด มอร์แดนท์ และได้ยกตัวอย่างงานวิจัยที่ศึกษาการใช้สีสกัดจากพืชใน การย้อมเนื้อเยื่อชนิดต่างๆ ซึ่งอาจนํามาใช้เป็นข้อมูลพื้นฐานและเป็นทางเลือกในการศึกษาและพัฒนาการย้อม สีเนื้อเยื่อจากสีที่สกัดจากพืช คําสําคัญ: สีย้อมจากพืช สีย้อมจากธรรมชาติ การย้อม เนื้อเยื่อสัตว์ #ผู้รับผิดชอบบทความ สัตวแพทย์มหานครสาร. 2557. 9(1): 63-78. E-mail address: [email protected] Araya Suebkhampet and Nattakarn Naimon / J. Mahanakorn Vet. Med. 2014. 9(1): 63-78. Using Dye Plant Extract for Histological Staining 1,# 2 Araya Suebkhampet and Nattakarn Naimon 1Department of Anatomy, Faculty of Veterinary Medicine, Mahanakorn University of Technology, Bangkok 10530, THAILAND 2Department of Veterinary Technology, Faculty of Veterinary Technology, Kasetsart University, Bangkok 10900, THAILAND Abstract: Nowadays, many and various kinds of dyes are used for histological staining due to the purpose of study. They are generally from both natural and synthetic origins. With the worldwide concern over the use of human health and eco-friendly materials, the use of natural dyes from plants has gain interested since most of them contain plenty of dye from their parts. They are also cheaper and more sustainable than the synthetic dyes. A brief review of plant dye classification based on their chemical structures of major dye pigments, methods of plant dye extraction, mordants and some of its application in diverse tissues are presented in this report. These would be helpful to further study and development of alternative tissue stains from plant natural dyes for tissue staining. Keywords: Plant dye, Natural dye, Staining, Animal histology #Corresponding author J. Mahanakorn Vet. Med. 2014. 9(1): 63-78. E-mail address: [email protected] Introduction are obtained from natural sources such as Histology is the study of cells and plants, insects, animals, clays and minerals tissues by using a light microscope. The (Carleton et al., 1976). Plants and insects ability to visualize or identify histological known to be used in histological staining for structures is frequently enhanced through animal tissues are Haematoxylon the use of histological stains. They give campechianum (logwood), from which contrast to the tissue as well as highlighting haematoxylin stain is obtained and particular features of interest. There are two Dactylopius cacti, from which carmine stain types of dyes that classified by their origins, is obtained (Egbujo et al., 2008), synthetic and natural dyes. The natural dyes respectively. The majority of natural dyes 64 Araya Suebkhampet and Nattakarn Naimon / J. Mahanakorn Vet. Med. 2014. 9(1): 63-78. are extracted from plant parts such as roots, Many dyes, techniques and procedures barks, leaves, berries and seeds and wood. are utilized to stain various types of animal The dyeing with natural dyes was one tissues. At present, haematoxylin and eosin of the oldest techniques practiced by the stain (H&E) is widely used as a light ancient people such as wall paintings in the microscopical stain in histology and caves or textile dyeing. The first use of dye histopathology. It is combination of natural in histology credited to Antonie van haematoxylin stain and the synthetic eosin Leeuwenhoek, the father of microbiology stain. Haematoxylin stains the cell nuclei who worked with saffron, a natural dye dark blue while eosin stains cell cytoplasm, extracted from saffron crocus. In the mid most connective tissue fibers and matrices in 1800s, amateur microscopists first used varying shades of pink and red. haematoxylin to stain cellular components Haematoxylin obtained from logwood tree (Titford M., 2005). Later scientists developed that is native to the Mexico and northern a wide range of haematoxylin staining Central America (Baker & Silverton, 1976), techniques. The systemically studied dye for but is now mainly cultivated in the West histology started in the second half of the Indies (Bancrot and Gamble, 2008). 19th century by Weigert C, J Gerlach, P Haematoxylin can be prepared in numerous Ehrlich and H Gierke. By then coloring ways and applied to tissues from different materials were mostly still natural origin like sites. It has to be oxidized to haematein carmine, cochineal, haematoxylin and indigo. before using (Culling, 1974). Haematein can Gradually the use of the natural dyes has be produced from haematoxylin in two decreased due to the introduction of ways: natural oxidation and chemical synthetic dyes (aniline dye from extract of oxidation. For natural oxidation, coal tar) which first invention by William haematoxylin is exposed to light and air Henry Perkin in 1856. They widely applied in (Ehrlich’s and Delafield’s haematoxylin many fields as food, cosmetic and textile solution), and for chemical oxidation, industries (Prabhu and Bhute, 2012). They sodium iodate (e.g. Mayer’s haematoxylin) or are rapid stain with vast range of new colors, mercuric oxide (e.g. Harris’s haematoxylin) easy to use and commercially available. are used. Natural oxidation of haematoxylin From then, the synthetic dyes together with seems to retain its staining ability longer those for histological staining have than that of the chemical oxidation. developed. However, it takes much longer time for preparation. Haematoxylin has also been 65 Araya Suebkhampet and Nattakarn Naimon / J. Mahanakorn Vet. Med. 2014. 9(1): 63-78. prepared synthetically (Morsingh and potential use in various purposes. It is Robinson, 1970) but the natural dye is interesting that over 2000 dyes are mostly used. Haematein is anionic charge, synthesized from various parts of plants, of having a poor affinity for tissues without the which only about 150 have been presence of mordant. A mordant is a commercially exploited (Gulrajani, 1992). substance used to set dyes on tissue They are used for applying to different sections. The common mordants for substrates such as textiles, paper, leather, haematoxylin are aluminium, iron and wood, hair etc. They are also widely used in tungsten. The dye-mordant complex cosmetic, food and pharmaceutical possesses a net positive charge and enables industries. A number of them own the to bind to anionic tissue regions including pharmaceutical properties and have been nuclear chromatin. Mordant is regularly used in traditional medicine. Curcumin, the included in the dyeing protocols when bright yellow dye of turmeric has been used natural dyes are used in order to fix or traditionally as a remedy for stomach and intensify the stains in cell or tissue staining. It liver ailments, as well as topically to heal is mainly a polyvalent metal ion that forms sores and revitalizes the skin (Chaturvedi, coordination complexes with certain dyes 2009). It has anti-oxidant, anti-inflammatory, which then attaches to the tissues anti-cancer and anti-septic properties (Llewellyn, 2005). Different kinds of (Chengaiah et al., 2010). Many other mordants give a different hue and stability common natural dyes such as lycopene (red of the dye in the tissues. dye) in tomatoes or lawsone (varies in color The use of natural dyes which are from orange to reddish brown) in henna are cheaper, eco-friendly and biodegradable has reported as antimicrobial agents (Siva, 2007; become a matter of significant important Habbal et al., 2011). due to the increased environmental In last 10 years, natural dyes have awareness and in order to avoid some been studied for their potential use in hazardous synthetic dyes to humans and various fields including histological staining. animals (Eom, et al., 2001). Several synthetic The present report describes the basic dyes (e.g. dyes with azo bonds nitro- or information about of plant dye classification amino-groups) cause allergic-like symptoms based on their chemical structures, methods or are carcinogenic (Ratna and Padhi, 2012). of the dye extraction, mordants and some of Therefore, alternative natural dyes have its application for tissue staining. These been currently more interested for their would be useful information to the 66 Araya Suebkhampet and Nattakarn Naimon / J. Mahanakorn Vet. Med. 2014. 9(1): 63-78. researchers who are searching for the new temperature and pH which depend on the natural dyes for tissue staining. properties of particular dye components (Prabhu and Bhute, 2012). After extraction, Classification of natural dyes from plants the extracts are generally filtered through Natural dyes can be classified in various filters such as cheesecloth, cotton different ways such as based on their wool or paper filter. The filtrates may
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