| Abstract [eng] |
Modification of Hematoxylin-Eosin Staining Method for Visualization of Elastic Fibres The topic of this Thesis has been chosen due to complicated diagnostics of intravenous tumor invasion, which is one of the tumors characterization features. In order to evaluate intravenous tumor invasion, visualization of blood vessels is necessary. The usual hematokxylin – eosin (H&E) method can only identify large blood vessels. Elastic fibers are not specifically distinguished by the H&E method, which requires additional staining methods. Considering these problems, the aim of this reaserch is to offer a modified H&E staining by adapting it to highlight elastic fibers for diagnostics of intravenous tumor invasion. Main tasks of the reaserch: analyze the significance of elastic fiber diagnostics; try out different combinations of H&E and elastic fibers staining techniques, selecting the most appropriate to vizualize intravenous invasion of tumor; apply proposed staining method for diagnostic work and compare results with H&E staining method. Literature analysis was first carried out: the diagnostic value of intravenous tumor invasion was analyzed, data about H&E and elastic fiber staining methods were collected. Based on the information gathered, 14 experiments were performed. The modification of H&E method (H&E_ORS) was developed during the experiments, which allows to highlight the elastic fibers in red color, nuclei and cytoplasm - blue, surrounding tissues – pale green, red blood cells - bright green color. This method cannot replace the usual H&E method but supplements it with new information and diagnostic capabilities. Results obtained: elastic fibers - dark red color, nuclei and cytoplasm - blue, surrounding tissue structures – pale green, red blood cells - bright green. This modification was used for the research. 30 cases were selected for the research: 10 lung and 20 rectum colon fragments with tumor. The test tissues were prepared using the usual FFPI technique. Tumor sections of all subjects were additionally stained with H&E_ORS modification. Pathologists have evaluated the possibilities of H&E and H&E_ORS methods to highlight the nuclei, elastic fibers and intravenous tumor invasion. The study found the superiority of the H&E method in nuclear visualization: in 96.67% of cases, the nuclei were well highlighted with only 36.67% by the H&E_ORS method. Reverse result observed in the evaluation of elastic fibers: 66.67% of cases in the H&E elastic fibers are not identified, good visualization is observed in 76.67% of cases by H&E_ORS method. The results of the intravenous invasion of the tumor have shown the limitations of the informative approach of the H&E method. After evaluating H&E preparations, positive intravenous tumor invasion was observed in only 13.33% of cases, with additional evaluation of HE_ORS preparations in 46.67% of cases. The results show the diagnostic value of the HE_ORS method in diagnostics of intravenous tumor invasion. |
