Comparative Study of Microwave Oven-Assisted Tissue-Processing with Xylene and Xylene Substitutes on Morphological Quality of Tissue Sections

2021 ◽  
Vol 20 (4) ◽  
Author(s):  
Sri Darsheny ◽  
Saint Nway Aye ◽  
Purushotham Krishnappa ◽  
Rashindra Ravindran

Background: Over several years, xylene has been traditionally utilised as the clearing agent of choice in tissue-processing due to effectiveness in rapidly clearing tissue, facilitating the paraffin infiltration process. However, xylene use adversely impacts the health of personnel with long term exposure due to toxicity. In order to overcome these effects and replace it with a safer alternative agent, the present study aims to compare quality of tissue sections processed using an isopropanol and mineral oil mixture and propylene glycol methyl ether (PGME) as xylene substitutes.   Methods: Rat skeletal muscle tissue samples (n=20) were prepared for each processing protocol with xylene substitutes. Tissue specimens were processed according to the proposed microwave protocol. The clearing steps were performed using isopropanol and mineral oil mixture, and PGME, replacing xylene. From each paraffin-embedded block, one section of 4-5µm thickness tissue was obtained and conventionally-stained with Haematoxylin and Eosin (H&E). The histological sections were microscopically assessed and scored by a pathologist. A qualitative analysis was performed with the results obtained.   Results: The overall score obtained for xylene processed tissue was 100% with a score of 2 for all the 3 parameters assessed. However, the outcome for tissue processed with isopropanol and mineral oil mixture was 28.6% unsatisfactory, 28.6% satisfactory and 42.8% good. In PGME-treated tissues, 14.3% were unsatisfactory sections, 71.4% were satisfactory and 14.3% produced good quality sections. Overall, tissues processed using both substitutes exhibited sufficient staining quality in terms of the aforementioned parameters as compared to xylene-processed tissues, though significant difference in scores were observed.   Conclusion: Despite several challenges faced in the study, isopropanol and mineral oil mixture and PGME can be suggested as alternative clearing agents to xylene, provided having access to a more sophisticated microwave oven with precise temperature control for complete tissue-processing.


2002 ◽  
Vol 126 (5) ◽  
pp. 583-590 ◽  
Author(s):  
Azorides R. Morales ◽  
Harold Essenfeld ◽  
Ervin Essenfeld ◽  
Maria Carmen Duboue ◽  
Vladimir Vincek ◽  
...  

Abstract Context.—Current conventional tissue-processing methods employ fixation of tissues with neutral buffered formalin, dehydration with alcohol, and clearing with xylene before paraffin impregnation. Because the time required for this procedure is usually 8 hours or longer, it is customary to process tissues in automated instruments throughout the night. Although this time-honored method continues to serve histology laboratories well, it has a number of shortcomings, such as a 1-day delay of diagnosis, the need to batch specimens, the relatively large volumes and toxicity of reagents used, and the extent of RNA degradation. Objective.—To describe a rapid new method of tissue processing using a continuous-throughput technique. Design.—We used a combination of common histologic reagents, excluding formalin and xylene, as well as microwave energy, to develop a rapid processing method. The effect of this method on the quality of histomorphology, histochemistry, immunohistochemistry, and RNA content of processed tissue was compared with that of adjacent tissue sections processed by the conventional processing technique. We also assessed the impact of this rapid processing system on our practice by comparing the turnaround times of surgical pathology reports before and after its implementation. Results.—The new processing method permitted preparation of paraffin blocks from fresh or prefixed tissue in about 1 hour. The procedure allowed continuous flow of specimens at 15-minute intervals. It eliminated the use of formalin and xylene in the processing and used considerably lower volumes of other chemical reagents. Histomorphologic, histochemical, and immunohistochemical results were comparable to the parallel sections prepared by the conventional method. The new technique, however, preserved higher quality RNA. Use of the new methodology led to the diagnosis and reporting of more than one third of surgical pathology specimens on the same day that they were received, as compared to 1% of same-day reporting before the implementation of the rapid processing system. Conclusion.—The quality of hematoxylin-eosin, histochemical, and immunohistochemical tissue sections provided by the new system is comparable to that obtained following the conventional processing method. The new system preserves RNA better than the conventional method. It also shortens the processing time to about 1 hour from the receipt of fresh or prefixed tissue, eliminates the need for formalin and xylene, and reduces the volume of other chemicals. Most importantly, it impacts overall patient management by allowing for considerably shorter turnaround times for completion of surgical pathology reports.


2021 ◽  
pp. 003232172110261
Author(s):  
Richard Nadeau ◽  
Jean-François Daoust ◽  
Ruth Dassonneville

Citizens who voted for a party that won the election are more satisfied with democracy than those who did not. This winner–loser gap has recently been found to vary with the quality of electoral democracy: the higher the quality of democracy, the smaller the gap. However, we do not know what drives this relationship. Is it driven by losers, winners, or both? And Why? Linking our work to the literature on motivated reasoning and macro salience and benefiting from the Comparative Study of Electoral Systems project—covering 163 elections in 51 countries between 1996 and 2018, our results show that the narrower winner–loser gap in well-established electoral democracies is not only a result of losers being more satisfied with democracy, but also of winners being less satisfied with their victory. Our findings carry important implications since a narrow winner–loser gap appears as a key feature of healthy democratic systems.


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