dehydration of human endometrial tissue for sectioning

Análisis de imagen de tejido endometrial teñido inmunohistoquímicamente

Recurrent miscarriage (RM) is the loss of two or more consecutive pregnancies and is a complex disease drawing attention from clinicians in recent years.The incidence rate of RM in women of childbearing age is 1%-5% 1. Results of previous studies show that immune factors are closely associated with the pathogenesis of RM2,3,4,5. Maintaining immune homeostasis at the maternal-fetal interface is required for embryo implantation and development. Endometrial immune cells perform several regulatory roles to maintain this homeostasis, such as promoting trophoblast invasion, remodeling spiral arteries, and contributing to placenta development6,7,8,9.
Aberrant endometrial immune cells in women with RM have previously been reported. Results show a close association between the high density of uterine natural killer cells (uNKs) and the occurrence of RM10,11,12. An increased number of macrophages has been reported in the endometrium of women with RM, compared with those who had a live birth13. Regulatory T cells (Treg) play a role in maternal immune tolerance toward the embryo, and their level and function are decreased in the decidua of RM patients14. Cytotoxicity T cells (CTL) and dendritic cells (DCs) also play a role in the immune regulation of pregnancy15,16. Therefore, a comprehensive quantitative analysis of local endometrial immune cells during the mid-luteal phase could help to better understand the pathogenesis of RM. Some current methods for quantitative analysis of endometrial immune cells use flow cytometry which can accurately label immune cells with multiple markers17,18. However, clinical application of flow cytometry is limited because it can only be performed on fresh tissue. Obtaining fresh tissue is only feasible when a large volume of excess tumor is available, a rare occurrence for endometrium. Immunohistochemistry can observe tissue morphology well in situ and can also label various immune cells, while traditional immunohistochemical techniques cannot perform quantitative analysis of immune cells.
Compared to conventional immunohistochemistry experiments, quantitative immunohistochemical analysis of immune cells in the endometrium has important clinical significance. IHC intensity scoring is usually ranked on a four-point scale or strong and weak in pathological diagnostics and research19,20,21. However, this semi-quantitative technique is subjective, highly inaccurate, and demonstrates significant intra-observer and inter-observer variability22. One possible solution is the application of machine learning, which is valuable indigital image analysis23,24. By providing quantitative measurements, this approach enables a more precise assessment of immune cell infiltration, distribution, and density within the uterine tissue. This quantitative information can help to elucidate the dynamic changes in immune cell populations during the menstrual cycle and in various pathological conditions. Overall, the ability to quantitatively analyze immune cells in the endometrium through immunohistochemistry offers valuable insights into the immune microenvironment of the uterus.
Therefore, the protocol aimed to developed and validated a digital immunohistochemistry image analysis platform to quantitatively analyze endometrial immune cells including uNK cells, Tregs, macrophages, DCs and cytotoxic T cells during the mid-luteal phase in RM patients.

Autor destacado: Avance de la inmunología reproductiva con un protocolo para la evaluación cuantitativa de células inmunitarias endometriales 

Plataforma para la detección cuantitativa de células inmunes endometriales basada en inmunohistoquímica y análisis de imágenes digitales

Watch this Scientific Journal Video about Dehydration of Human Endometrial Tissue for Sectioning at JoVE.com

Deshidratación del tejido endometrial humano para seccionamiento

Para empezar, utiliza unas pinzas para transferir el tejido endometrial de la formalina al papel de inclusión. Con una regla, mide las dimensiones del tejido endometrial. Ahora envuelva el pañuelo en el papel de incrustación y colóquelo en un casete de incrustación.Transfiera el casete a la canasta de pañuelos desechables para deshidratarlo. Para deshidratar el tejido endometrial, coloque la canasta de tejido en la cámara de reacción del deshidratador e inicie el procedimiento de deshidratación de tejido de rutina. Una vez completada la deshidratación, abra la cámara de reacción del deshidratador y retire la canasta de pañuelos.

dehydration-of-human-endometrial-tissue-for-sectioning

Research

JoVE Journal

Biology

Dehydration of Human Endometrial Tissue for Sectioning

89 vistas. Para empezar, utiliza unas pinzas para transferir el tejido endometrial de la formalina al papel de inclusión. Con una regla, mide las dimensiones del tejido endometrial. Ahora envuelva el pañuelo en el papel de incrustación y colóquelo en un casete de incrustación.Transfiera el casete a la canasta de pañuelos desechables para deshidratarlo. Para deshidratar el tejido endometrial, coloque la canasta de tejido en la cámara de reacción del deshidratador e inicie el procedimiento ...

Video: Deshidratación del tejido endometrial humano para seccionamiento

Platform for Quantitative Detection of Endometrial Immune Cells Based on Immunohistochemistry and Digital Image Analysis

To evaluate the endometrial immune microenvironment of patients with recurrent miscarriage (RM), a digital immunohistochemistry image analysis platform was developed and validated to quantitatively analyze endometrial immune cells during the mid-luteal phase. All endometrium samples were collected during the mid-luteal phase of the menstrual cycle. Paraffin-embedded endometrial tissues were sectioned into 4 &#181;m thick slides, and immunohistochemistry (IHC)staining was carried out for detecting endometrial immune cells, including CD56+ uNK cells, Foxp3+ Tregs, CD163+ M2 macrophages, CD1a+ DCs, and CD8+ T cells. The panoramic slides were scanned using a digital slide scanner and a commercial image analysis system was used for quantitative analysis. The percentage of endometrial immune cells was calculated by dividing the number of immune cells in the total endometrial cells. Using the commercial image analysis system, quantitative evaluation of endometrial immune cells, which are difficult or impossible to analyze with conventional image analysis, could be easily, and accurately analyzed. This methodology can be applied to quantitatively characterize the endometrium microenvironment, including interaction between immune cells, and its heterogeneity for different reproductive failure patients. The platform for quantitative evaluation of endometrial immune cells may be of important clinical significance for the diagnosis and treatment of RM patients.

Here, a digital immunohistochemistry image analysis platform was developed and validated to quantitatively analyze the endometrial immune cells of patients with recurrent miscarriages in the window of implantation.

To evaluate the endometrial immune microenvironment of patients with recurrent miscarriage (RM), a digital immunohistochemistry image analysis platform ...