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Research Article Open access CC BY 4.0

Assessment of the Conventional versus Two Rapid Microwave Processing Methods Using the Phosphotungstic Acid Haematoxylin Technique

Tobias Peter Pwajok Choji, Anthony Ajuluchukwu Ngokere, Samuel Ifedioranma Ogenyi, Peterside Rinle Kumbish, Lovelyn Peter Unubi, Mada Umoru Alesa, Ponfa Nden Zhakom, Emmanuel Ishaku Vomangai, Gwom Irmiya Davou, Charity Ezikiel Marcus

Annual Research & Review in Biology · pp. 1–18 · Published 25 May 2016

10.9734/ARRB/2016/19219

Abstract

Aims: To process tissues using the rapid microwave techniques and check their reactivity with the phosphotungstic acid haematoxylin. To establish a very fast and cheap method of tissue processing and evaluate its effects on tissue morphology and dye uptake. To eliminate xylene from the processing schedule. To check for additional information that can be obtained from the phosphotungstic acid haematoxylin technique so as to encourage its use in the laboratory. Study Design: Harvesting and fixation of tissues. Grossing into triplicates, processing using three different techniques. Staining and grading of sections. Place and Duration of Study: Human samples were obtained from the Anatomy Department, Nnamdi Azikiwe University, Nnewi Campus while animal samples were obtained from the Animal experimental station, National veterinary Research institute, Vom, Nigeria. The work was done between August and December, 2014. Methodology: Heart, skin, brain, kidney, lungs, and liver tissues were harvested from an apparently healthy rabbit after sacrificing it scientifically. Ethical clearance as well as human cervix, fibroid, prostate and testis were obtained. All samples were fixed in 10% buffered formalin, cut into triplicates and processed using the conventional as well as two rapid microwave techniques respectively. Sections were made from each technique and stained simultaneously using the Phosphotungstic Acid Haematoxylin technique. The Masson Trichrome methos as well as the Haematoxylin and Eosin Techniques were also used to assess morphology They were each evaluated for quality of block, ribboning, nuclear, cytoplasmic and extracellular components, collagen and muscle fibre. Section thickness, uniformity and staining intensities were all checked. Results: Processing cycle as well as quantity of reagents used was drastically reduced in the microwave techniques. Paraffin blocks obtained, ribbonning as well as flattening on 20% alcohol were comparable in the three techniques. Tissue morphology, integrity of tissue as well as tissue histochemistry to the phosphotungstic acid haematoxylin were comparable among the three techniques. The stroma of the cervix is better retained in the microwave with vacuum technique. Conclusion: Samples received in histology laboratory can be processed and slides produced within the same day without compromising tissue morphology or histochemistry. Xylene can be eliminated from the paraffin wax processing cycle with quality paraffin block and ribbon production. The cost and duration of tissue processing can be drastically reduced with the production of quality sections. This work can be replicated using other histochemical techniques to give a wider application.

Paraffin wax method processing cycle reagent toxicity microwave isopropanol histochemistry phosphotungstic acid haematoxylin.

References (15)

  1. 1 Xylene: An overview of its health hazards and preventive measures [DOI]
  2. 2 Xylene-Free Method for Histological Preparation: A Multicentre Evaluation [DOI]
  3. 3 Current Concepts of Enzyme Histochemistry in Modern Pathology [DOI]
  4. 4 Microwave irradiation as a form of fixation for light and electron microscopy [DOI]
  5. 5 Microwave‐stimulated diffusion for fast processing of tissue: reduced dehydrating, clearing, and impregnating times [DOI]
  6. 6 Continuous-Specimen-Flow, High-Throughput, 1-Hour Tissue Processing [DOI]
  7. 7 Pathologic detection of early myocardial infarction: a critical review of the evolution and usefulness of modern techniques.
  8. 8 Evaluation of Ethanol-based Fixatives as a Substitute for Formalin in Diagnostic Clinical Laboratories [DOI]
  9. 9 Ultra-rapid microwave-stimulated tissue processing with a modified protocol incorporating microwave fixation [DOI]
  10. 10 Microwave histoprocessing versus conventional histoprocessing [DOI]
  11. 11 Staining reactions of microwave processed tissues compared with conventional paraffin wax processed tissues
  12. 12 Microwaves for microscopy [DOI]
  13. 13 Microwave technique in histopathology and its comparison with the conventional technique.
  14. 14 The two-step vacuum-microwave method for histoprocessing.
  15. 15 Evaluation of Ethanol-based Fixatives as a Substitute for Formalin in Diagnostic Clinical Laboratories [DOI]

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