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Influence of fixation time and fixation type on immunohistochemical analysis in the detection of stem cells in endometrial tissue.
Macháčová, Dominika ; Nachtigal, Petr (advisor) ; Němečková, Ivana (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Title of Diploma Thesis: Influence of fixation time and type of fixative solution on immunohistochemical analysis in the detection of stem cells in endometrial tissue Author: Bc. Dominika Macháčová Supervisor of Diploma Thesis: prof. PharmDr. Petr Nachtigal, Ph.D. Backgroud: The aim of this diploma thesis was to investigate the effect of the type of fixative solution and the length of fixation on immunohistochemistry. Materials and methods: A total of five porcine and five bovine endometria obtained from hysterectomy specimens were included in this study. Immunohistochemical staining with progesterone receptors and SOX2 in endometrium fixed in five fixative solutions (Formol, Antigenfix, Greenfix, Bouin, Methacarn) were performed after 1,5; 8,5; 15,5; 29,5; 64,5 and 189,5 days of fixation. An automated immunostainer Ventana Benchmark Ultra was used for all immunohistochemical analysis. Results: Expression of progesterone receptors was demonstrated in 92-87 % of glandular cells after 1,5-64,5 days and in 59 % of cells after 189,5 days of formalin fixation. The intensity of staining was strong, but after 189,5 days it was evaluated as being weak. In Antigenfix-fixed tissues, strong progesterone...
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The use of immunohistochemistry for the examination of mammary gland biopsy.
MACHÁČOVÁ, Dominika
Background: Tumours of the breast are the most common tumours in the female population of the Czech Republic. Improvements in early diagnosis and therapy have led to gradual reduction of mortality with an increasing incidence of this disease. Aims: The aim of this study was to find out the significance of imunohistochemistry in diagnostics of tumours of the breast. Materials and methods: A total of 22 cases of malignant neoplasm of the breast were included in our study. Twenty two core-cut biopsies and twenty one resected specimens were eximined. Imunohistochemical staining with a total of 5 markers was performed - oestrogen and progesterone receptors, HER2, Ki-67, E-cadherin. In 9 cases, imunohistochemical examination in the resection specimens was also performed. The results of manual imunohistochemical staining and staining performed on autostainer Ventana Benchmark were compared. Results: The most frequent histologic subtype was invasive carcinoma of no special type in fourteen of cases, followed by four cases of invasive lobular carcinoma, two cases of atypical medullary carcinoma, one case of mixed (invasive carcinoma of no special type and invasive lobular carcinoma) carcinoma and one case of metaplastic carcinoma. In one case, a different diagnosis was made - invasive carcinoma of no special type was diagnosed in the core-cut biopsy while metaplastic carcinoma was diagnosed in the resected specimen. The expression of both oestrogen and progesterone receptors was variable. It ranged between 10-100 % (on average 81,9 % and 38,8 % respectively). Oestrogen receptors were found in 18 of cases while progesterone receptors were found in 16 of cases. The Ki-67 expression ranged between 2 and 40 % (on average 16,5 %); higher Ki-67 values ( 20 %) were found in tumours with loss of oestrogen and/or progesterone receptor expression and in poorly differentiated tumours. None of the cases was determinated as HER2 positive; a score 0 was used in 12 of the cases and a score 1+ in 10 of the cases. Conclusion: When comparing manual imunohistochemical staining staining with staining performed on the autostainer Ventana Benchmark very similar results were obtained.
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