A simple method for the prevention of endometrial autolysis in hysterectomy specimens

Houghton, Joseph; Roddy, Sarah; Carroll, Sarah B.; McCluggage, W. Glenn

doi:10.1136/jcp.2003.012898

Cited by 14 publications

(5 citation statements)

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“…If the uteri could not be sectioned within 15 minutes, before placing them in 10% formalin, they were sliced coronally through the endometrium to minimize endometrial autolysis that could adversely affect subsequent immunostaining (23,24). Sections of endometrium were obtained from six different regions of the endometrial cavity: fundus, left cornu, right cornu, anterior body, posterior body, and LUS.…”

Section: Methodsmentioning

confidence: 99%

Quantification of nucleolar channel systems: uniform presence throughout the upper endometrial cavity

Szmyga

Rybak

Nejat

et al. 2013

Fertility and Sterility

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Objective To determine the prevalence of nucleolar channel systems (NCSs) by uterine region applying continuous quantification. Design Prospective clinical study. Setting Tertiary care academic medical center. Patients 42 naturally cycling women who underwent hysterectomy for benign indications. Intervention NCS presence was quantified by a novel method in six uterine regions, fundus, left cornu, right cornu, anterior body, posterior body, and lower uterine segment (LUS), using indirect immunofluorescence. Main Outcome Measures Percent of endometrial epithelial cells (EECs) with NCSs per uterine region. Results NCS quantification was observer-independent (intraclass correlation coefficient [ICC] = 0.96) and its intra-sample variability low (coefficient of variability [CV] = 0.06). 11/42 hysterectomy specimens were midluteal, 10 of which were analyzable with 9 containing over 5% EECs with NCSs in at least one region. The percent of EECs with NCSs varied significantly between the lower uterine segment (6.1%; IQR = 3.0-9.9) and the upper five regions (16.9%; IQR = 12.7-23.4) with fewer NCSs in the basal layer of the endometrium (17% +/−6%) versus the middle (46% +/−9%) and luminal layers (38% +/−9%) of all six regions). Conclusions NCS quantification during the midluteal phase demonstrates uniform presence throughout the endometrial cavity, excluding the LUS, with a preference for the functional, luminal layers. Our quantitative NCS evaluation provides a benchmark for future studies and further supports NCS presence as a potential marker for the window of implantation.

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Section: Methodsmentioning

confidence: 99%

Quantification of nucleolar channel systems: uniform presence throughout the upper endometrial cavity

Szmyga

Rybak

Nejat

et al. 2013

Fertility and Sterility

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“…Formalin fixation is the primary and universal method of tissue processing for light microscopy and archival preservation because it prevents tissue autolysis and preserves histological and morphological architecture . Some tissue repositories (e.g., brain banks) contain thousands of formalin-preserved or formalin-fixed paraffin-embedded specimens representing an invaluable source for histopathological examination and retrospective clinical studies .…”

mentioning

confidence: 99%

Antigen-Epitope Retrieval To Facilitate Proteomic Analysis of Formalin-Fixed Archival Brain Tissue

et al. 2006

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Formalin is a routine fixative facilitating tissue preservation and histopathology. Proteomic techniques require freshly frozen specimens, which are often difficult to procure, and methods facilitating proteomic analysis of archival formalin-fixed brain tissue are lacking. We employed antigen-epitope-retrieval principles to facilitate proteomic analysis of brain tissue that had been fixed and stored in formalin for 3-7 years. Twenty-micrometer-thick cryopreserved OCT-embedded sections from inferior temporal cortex of human (7 years in formalin) or mouse brain specimens (3 years in formalin) were hematoxylin-/eosin-stained. Approximately 16-64-mm2 areas of the tissue sections were manually scraped off slides, or approximately 2 mm2 of human brain cortex was captured off membrane-coated slides using laser microdissection. Tissue was treated using various pH and temperature conditions prior to trypsin digestion and nano-LC-MS/MS. The largest number of proteins were retrieved by solubilization at pH 9 at 95 degrees C for 1 h; treatments at pH 4 or 6 at 25 or 65 degrees C were generally ineffective. Three-year formalin-fixed murine tissue did not yield more proteins compared to human tissue. Use of formalin-fixed tissue for proteomics is an invaluable tool for medical research. The combination of proteomics and microdissection enables selective enrichment and identification of novel, unique, or abundant proteins that may be important in pathogenesis.

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“…Although these investigators did not specifically test the effect of the fixation protocol on immunostain quality, we hypothesise this was a major player in the problems faced in the pilot phase of our project. Some authors have prevented these autolytic changes by injecting formalin through the cervical os to improve fixation while preventing uterine wall distortion [42].…”

Section: Fixation Quality and Mmr Immunostainingmentioning

confidence: 99%

Mismatch Repair Protein Expression Defects in Endometrioid Endometrial Adenocarcinoma

Brincat¹

2020

Preprint

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Objectives: Endometrioid endometrial carcinoma (EEC) is the sentinel cancer in over half female patients with heritable mismatch repair (MMR) mutations as part of Lynch syndrome. Immunohistochemical testing for tumoural MMR-protein expression is the primary screening test identifying cases potentially harbouring familial cancer syndrome-related mutations and is also a predictive biomarker for immune-checkpoint blockade response. Methods: Following Data Protection and Ethical clearance by the University of Malta, 200 EEC cases were retrospectively identified and categorized into three arms: 151 cases above age 50 at diagnosis, 49 cases at or below age 50 at diagnosis and 30 controls with benign endometrial tissue sampling. H&E case slides were re-examined by an independent pathologist to confirm the diagnosis and identify the block best representing the tumour. Four new slides per case were recut and immunohistochemistry performed for MLH1, PMS2, MSH2, and MSH6 proteins. Protein expression was analysed semiquantitatively using Allred scoring. Results: 31% of the overall EEC cases were deficient for one or more MMR-proteins. Dual loss of the MLH1-PMS2 protein heterodimer was the most common deficiency, occurring in 24.5% of cases. Loss of MSH2-MSH6 protein expression represented 3.2% of MMR-deficient cases. Well di ferentiated tumours had a 76.5% proficiency rate as opposed to grade 2/3 disease with 53.2% and 52.9% proficiency rate respectively. There was no significant di ference in MMR status when age 50 was used as a hypothetical testing threshold. A ter correcting for tumour grade, MLH1 and PMS2 expression was shown to be negatively correlated with age-at-diagnosis while MSH6 expression was positively correlated. Conclusion: Re lex MMR proficiency testing of all EEC cases is advisable, as using age 50 as a testing threshold would have missed 82.3% of MMR deficient cases. Prospective evidence is required to clarify the role semi-quantitative scoring plays in MMR status interpretation and patient management in the ever-evolving field of targeted therapeutics.

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A simple method for the prevention of endometrial autolysis in hysterectomy specimens

Cited by 14 publications

References 0 publications

Quantification of nucleolar channel systems: uniform presence throughout the upper endometrial cavity

Quantification of nucleolar channel systems: uniform presence throughout the upper endometrial cavity

Antigen-Epitope Retrieval To Facilitate Proteomic Analysis of Formalin-Fixed Archival Brain Tissue

Mismatch Repair Protein Expression Defects in Endometrioid Endometrial Adenocarcinoma

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