The aim of this study is to compare the efficacy of xylene-free hematoxylin and eosin (H and E) sections with conventional H and E sections. Individual sections cleared J Oral Maxillofac Pathol 2014;18:49‑53. Tissues were evaluated for eight parameters: sectioning, nuclear staining, cytoplasmic staining, overall cell morphology, clarity of staining, uniformity of staining, quality of immunohistochemistry (IHC), and cost. The hematoxylin and eosin stain (H&E) is the most widely used stain in histology and histopathology laboratories. Given the preservation of structure and excellent sectioning property, formalin-fixed paraffin-embedding method had an enormous potential for three-dimensional reconstruction of fine structures, based on the very thin histological sections and optical images. Furthermore, tissue processes that do not contain formaldehyde were found to be superior on preservation of nucleic acids. To assess the impact of new alternative solutions to formaldehyde and xylene on tissue processing, 13 different tissue processings were designed and performed on thirteen different tissues by using five different fixatives (formaldehyde, Glyo-Fixx®, FineFix®, Cell-block®, Green-Fix®) and four different clearing agents (xylene, Sub-X®, Bio-clear®, Shandon Xylene Substitute®). in its physical and clearing properties, which also maintained We prepared 30 paired soft tissue specimens for routine histopathological evaluation using conventional xylene and xylene-free methods to evaluate and compare their efficacy for fixation, processing, embedding, staining and turnaround time. UltraClear™ is more expensive than xylene. The study concluded that the vegetable oils could be used as substitutes for xylene and clove oil. There is only. Our study has several limitations. Azer Scientific's Everclear™ Xylene substitute is a safe alternative to xylene. The commonest clearing agent is xylene. Conventional xylene and xylene-free methods for routine histopathological preparation of tissue sections. A total of 100 paraffin embedded tissue blocks from different tissues were included. Our study presents a new deparaffinizing and hematoxylin and eosin (H and E) staining method that involves the use of easily available, nontoxic and eco-friendly liquid diluted dish washing soap (DWS) by completely eliminating expensive and hazardous xylene and alcohol from deparaffinizing and rehydration prior to staining, staining and from dehydration prior to mounting. Results: Results showed that all the four oils had ability to However, the hazards associated with exposure to xylene are of concern. Article Title: Alternative to xylene as a clearing agent in histopathology doi: 10.4103/JLP.JLP_111_17 Figure Lengend Snippet: Immunohistochemistry stain of gallbladder using smooth muscle actin marker at magnification ×40: (a) UltraClear processed, (b The effect of the alternative solutions to formaldehyde and xylene on tissue processing. effects on health of technicians and pathologists when exposed cytological xative: A pilot study. study compared xylene with three clearing agents, These investigators concluded that their clearing agents. On the accounts of health hazards and cost of conventional clearing agents, the study also recommended groundnut oil, coconut oil and palm kernel oil as alternative clearants in wood histological processes. ... Histosec® pastilles solidification point 56-58°C embedding agent for histology. eosin staining. We observed differences in tissue transparency, production of serial sections, and clarity of histological, staining between tissues processed using UltraClear™, or xylene. © 2008-2021 ResearchGate GmbH. Histochemical and immunohistochemical (IHC) staining results were compared with qualitative and quantitative data obtained by a computer program, respectively. and aromatic hydrocarbons, and various types of oils. principle of fluid dynamics in rapid clearing of tissues by using effect particularly those fixed in non-protein coagulant fixatives. Two smears were collected from each subject. Wajid Sermadi Department of Dentistry, Chamrajnagar Institute of Medical Sciences, Chamrajnagar, India. Cost-benefit balance between safety of laboratory workers, good quality staining, and cost-effective strategy needs to be further studied. It is a flammable liquid that requires utmost Score ≤2 was considered inadequate for diagnosis while scores 3–5 were considered adequate for diagnosis. UltraClear™ is less toxic, less flammable, friendlier to the environment, and easy to handle, but it is two times expensive than xylene. Therefore, the present review aims to briefly describe the xylene and its noxious effects and other safer compounds which may be an alternative to xylene. (2013) Comparison of conventional tissue processing with microwave processing using commercially available and domestic microwaves. Abstract A simple and effective method for the substitution of mineral oil for xylene in tissue processing is described. An alternative histoprocessing, program was set for particularly fatty tissues [, an embedding machine (HistoStar™; Thermo Fisher, All slides were stained with hematoxylin and, Prisma; Sakura, Nagano, Japan) to evaluate overall, tissue morphology. Results: Adequate nuclear staining was noted in 94% in group A and 96% in group B, -adequate cytoplasmic staining in 92% in group A and 86% in group B, clarity in 94% of group A and 96% of group B sections, uniform staining in 92% of group A and 80% of group B sections, crisp stain in 96% of group A and 88% of group B sections, and 94% of group A sections stained adequately for diagnosis as compared with 90% in group B sections. Slides were scored for parameters: nuclear, cytoplasmic, clarity, uniformity, and crispness of staining. In this study stability and longetivity Half of the dehydrated tissues were cleared in three, changes of xylene as follows: xylene one and two for 1 h, and each and xylene three for 2 h. The other group was, cleared in UltraClear™. were easy to cut and worked well for both hematoxylin and eosin and IHC staining. CONCLUSION: UltraClear is less toxic, less flammable, friendlier to the environment, and easy to handle, but it is two times expensive than xylene. We used 230 formalin-xed, parafn-embedded, Markers(Dako,Denmark) Antigenretrieval(PTLink,Dako,Denmark), =115 for each group. The choice of a clearing agent depends upon the type of tissue processor used, the processing conditions such as temperature, safety factors and cost. CitriSolv™ is a d-Limonene-based solvent and clearing agent that can be used as a safe alternative to xylene and ethyl acetateVWR offers a broad line of reagent droppers, diagnostic kits, and antigens and antisera, for aid in identifying a variety of microscopic organisms. Key message staining quality and a comparison was done between them. rapidly and renders transparency, facilitating clearing endpoint As previous research studies have shown the effectiveness of different vegetable oils as clearants, this study was designed to evaluate the efficacy of coconut oil. oils such as Carrot oil, Olive oil, Pine oil, Rose oil and xylene and Toluene: It is also a good clearing agent but less commonly used in histopathology laboratory. Group D evaluates UltraClear™ as both dewaxing and clearing agents prior to coverslipping. Groups B, C, and D showed 79% (P = 0.054), 83% (P = 0.221), and 80% (P = 0.079) adequacy when compared with Group A (89%), respectively. Sections processed using UltraClear™ were easy to cut (81.7%) as were xylene processed sections (96.5%). In this study, we introduced a novel, non-toxic xylene substitute (SBO), which was generated through a mixture of 86% of white oil No.2 and 14% of N-heptane. Materials and Methods Group C sections stained adequate for diagnosis (93.3%) followed by Group B (88.7%) and Group A (78%; P < 0.05). Despite the fact that there is a difference in the, percentages between UltraClear™ and xylene processed, tissues, histologists and pathologists in our institution. RESULTS the organic solvents and paraffin wax. UltraClear™ is a promising dewaxing agent. transferred into hot-air oven. is used as a solvent in the rubber, printing, The primary problem with using xylene is its, toxicity; long‑term exposure is particularly, injurious to health. to longer duration. Results: Results showed that all the four oils had ability to On exposure the vapours are rapidly absorbed through the lungs and the slowly through the skin. Clearing is an essential step in processing tissue for light microscopy. as clearing agents. Biotech Histochem 2012;87:464‑7. CONCLUSION Group D evaluates UltraClear™ as both dewaxing and clearing agents prior to coverslipping. According to Bernoulli’s principle of fluid dynamics, to decrease rapid clearing hot-air oven technique was used. Others have investigated alternative clearing agents such, as a novel, nontoxic xylene substitute (SBO). The slides were washed in tap water for about 30 s, following which they were subjected to the conventional Papanicolaou staining procedure. 10.4103/JLP.JLP_111_17 Chandraker R, C.Rathod V, K. Chandraker N, Pundir S, Dixit S, Desai V. Comparison Between Xylene And Coconut Oil In The P-value obtained was 0.47 and the data were statistically insignificant. A comparative study to evaluate liquid dish washing soap as an alternative to xylene and alcohol in deparaffinization and hematoxylin and eosin staining. Toluene is better at preserving tissue structure and is more tolerant of small amounts of water left behind in the tissues than xylene. to longer duration. For each Group of A, B, C, and D, 100 slides were cut from xylene-processed blocks. Under a light microscope, the results showed that sections cleared in the vegetable oils did not show any sign of distortion as details of wood features were distinct and clear after processing in the vegetable oils.
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