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Thermal Imaging for Arthritis Evaluation in a Small Animal Model
Published in U. Snekhalatha, K. Palani Thanaraj, Kurt Ammer, Artificial Intelligence-Based Infrared Thermal Image Processing and Its Applications, 2023
U. Snekhalatha, K. Palani Thanaraj, Kurt Ammer
A specimen of the RA affected joint is obtained through closed needle biopsy or arthroscopic guided biopsy. Then, it is stained with hematoxylin and eosin. The histological evaluation of synovial tissue inflammation in RA confirms the hyperplasia in the lining layer, cellular infiltration with lymphocytes, macrophages, granulomas, pannus formation, and plasma cells (Snekhalatha et al., 2012). In the Wistar rat-RA model, to confirm the pathological conditions, a histology study on rats was carried out by sacrificing on the 30th day and dissecting their forepaw and hind paw.
The Medicolegal Autopsy
Published in Kevin L. Erskine, Erica J. Armstrong, Water-Related Death Investigation, 2021
All major organs and glands are individually sectioned multiple times using a scalpel to look for structural abnormalities, disease, and injury. A small tissue sample of each major organ is taken and submitted in a fixative solution prior to histological preparation and later examined under the microscope by the forensic pathologist (Figure 7.51). This histological preparation involves standard application of stains called hematoxylin and eosin to each section of tissue in order to bring out the unique characteristics of the tissue and cellular structure (Figure 7.52). The number of samples taken for any given case is dictated by the complexity of the autopsy and the need to adequately characterize the extent of injury or disease. A typical autopsy may include sampling of the heart, lungs, liver, kidneys, and brain, more or less, depending on the case. In certain cases, the pathologist may opt not to submit tissue samples for microscopic examination. Sampling with long-term retention of small portions of tissue in a stock jar containing formalin fixative is routine. These reserved tissues are kept for a pre-defined and often limited time period according to the protocol of the office and is done in the event a future need for a microscopic examination of those tissues arises.
Laboratory Procedures and Management
Published in Jeremy R. Jass, Understanding Pathology, 2020
The next stage involves removing the wax and staining the sections. The slides are loaded into racks and placed in xylene to dissolve the wax. This is followed by 100%, 90% and 70% alcohol and finally back to water, whereby the sections become completely transparent. In order to be studied by microscopic examination, they are dyed with chemicals, usually haematoxylin, which stains nuclei blue (due to the presence of acidic DNA) and eosin which imparts a pink colour to the cytoplasm (H&E sections). The sections are then dehydrated and returned to xylene before being covered with a glass cover slip with the aid of runny and transparent mounting medium that slowly hardens. A senior technologist checks the final quality of the slides and then they are passed to the anatomical pathologist for reporting.
Hydroxysafflor yellow A inhibits endothelial cell ferroptosis in diabetic atherosclerosis mice by regulating miR-429/SLC7A11
Published in Pharmaceutical Biology, 2023
Jianjie Rong, Chuanyong Li, Qiang Zhang, Guangfeng Zheng, Weijian Fan, Zhichang Pan, Shuming Shi
The expression levels and distribution of the AS marker proteins intercellular adhesion molecule (ICAM) and vascular cell adhesion molecule (VCAM) in the aortic tissues of each group (n = 3) were measured using immunohistochemistry. After deparaffinization, the paraffin sections were incubated with 3% H2O2 for 20 min. Tissue antigen retrieval was performed at 121 °C for 20 min using citrate buffer (pH = 6.0). Then, 1% bovine serum albumin was added for 20 min. Rabbit anti-mouse ICAM (cat no. ab222736, Abcam) and VCAM (cat no. ab134047, Abcam) antibodies were added dropwise, and the tissues were incubated at 4 °C for 16 h. After washing away the unbound primary antibody, goat anti-rabbit IgG-HRP antibody (cat no. ab6721, Abcam) was added dropwise to the tissues and incubated at 37 °C for 1 h. 3,3’-Diaminobenzidine developer solution was used for color development. Hematoxylin was used to stain the nuclei. After dehydration with alcohol, the sections were mounted with neutral gum after the xylene became transparent. The results were photographed under an optical microscope (Leica). Statistical quantification of the positive densitometric values was performed using Image-Pro Plus 6.0.
Suberosin Alleviates Sepsis-Induced Lung Injury in A Rat Model of Cecal Ligation and Puncture
Published in Journal of Investigative Surgery, 2023
Sevgi Karabulut Uzunçakmak, Zekai Halıcı, Songül Karakaya, Zerrin Kutlu, Yavuz Selim Sağlam, İsmail Bolat, Pelin Aydın, Ceyda Sibel Kılıç
For immunoperoxidase examination, sections prepared on adhesive (poly-L-lysin) slides were treated with xylol and alcohol series. After being washed with PBS, these sections were incubated with 3% H2O2 for 10 min to inactivate the endogenous peroxidase. In order to collect antigens from tissues, samples were treated with antigen retrieval solution for 2 × 5 min at 500 W in a microwave oven and then left to cool. Tissues were incubated with IL-1β and TNF-α antibodies (Catalog Nos. sc-52012 and sc-52746, Santa Cruz Biotechnology, Santa Cruz, CA) at 37 °C for 30 min. Immunohistochemical (IHC) analysis was performed in accordance with the manufacturer’s protocol (Abcam HRP/DAB Detection IHC Kit) and 3.3′-diaminobenzidine (DAB) was used as the chromogen. Hematoxylin staining was applied. Sections were evaluated according to immunopositivity as none (−), mild (+), moderate (++), severe (+++), or very severe (++++) [8].
Myeloid-derived suppressor cells prevent disruption of the gut barrier, preserve microbiota composition, and potentiate immunoregulatory pathways in a rat model of experimental autoimmune encephalomyelitis
Published in Gut Microbes, 2022
Dušan Radojević, Marina Bekić, Alisa Gruden-Movsesijan, Nataša Ilić, Miroslav Dinić, Aleksandar Bisenić, Nataša Golić, Dragana Vučević, Jelena Đokić, Sergej Tomić
To evaluate cellular infiltrates in the spinal cords of animals with EAE, the spinal cords were isolated carefully from euthanized animals at the peak of the disease (15th-day post-immunization), and the lumbar part of the spinal cord was fixed in 4% paraformaldehyde for two weeks. Tissues were dehydrated in a series of ethanol (70%, 96%, 96%, and 100%) and xylol (33% in ethanol, 66%, 100%, and 100%) solutions, and then paraffin-embedded using the Tissue Embedding Center EC 350 (Especialidades Médicas Myr, Spain). Cross-sections 5–7 µm thick) were made using a manual microtome, and the slides were rehydrated in dilutions of xylol in PBS (100%, 96%, and 70%) prior to staining. Hematoxylin and eosin staining was performed using the corresponding dyes purchased from Sigma Aldrich, embedded in Canada balsam, and analyzed by light microscopy.