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Tissue and Molecular Diagnosis
Published in Professor Sir Norman Williams, Professor P. Ronan O’Connell, Professor Andrew W. McCaskie, Bailey & Love's Short Practice of Surgery, 2018
Professor Sir Norman Williams, Professor P. Ronan O’Connell, Professor Andrew W. McCaskie
A ‘special stain’ is a stain that is not routine, i.e. not an H&E stain. Immunohistochemical stains are conventionally excluded from this category. Some special stains demonstrate normal substances in increased quantities or in abnormal locations. The periodic acid-Schiff (PAS) stain demonstrates both glycogen and mucin, whereas a diastase PAS (D-PAS) stain demonstrates mucin, e.g. in an adenocarcinoma. Perls Prussian blue stain demonstrates iron accumulation (Figure16.23), e.g. in haemochromatosis. A reticulin stain helps to demonstrate fibrosis (Figure16.24). Elastic stains also show fibrosis and can highlight blood vessels by outlining their elastic laminae. Special stains can also reveal the accumulation of abnormal substances, e.g. a Congo red stain for amyloid.
Hematopoietic System
Published in Pritam S. Sahota, James A. Popp, Jerry F. Hardisty, Chirukandath Gopinath, Page R. Bouchard, Toxicologic Pathology, 2018
Kristin Henson, Tanasa Osborne, Gregory S. Travlos
Cytologic changes observed with dyserythropoiesis include abnormal nuclear shapes (peanut-shaped nuclei), asymmetric binucleation, premature pyknosis, nuclear fragmentation, nuclear to cytoplasmic maturation asynchrony (immature nuclei with hemoglobinized cytoplasm), maturation arrest, and iron-positive basophilic stippling (Harvey 2001). Erythrodysplasia is often associated with megaloblastic change or macrocytic response due to interference with DNA synthesis. For example, folate or cobalamin deficiencies, or administration of compounds that inhibit folate absorption from the intestines (ethanol, barbiturates, diphenylhydantoin), or inhibit cellular uptake of folate (methotrexate), result in megaloblastic anemia, with ineffective erythropoiesis due to inhibition of DNA synthesis and arrest of nuclear maturation and cell division (MacKenzie and Eustis 1990; Rebar 1993). Erythroid dysplastic changes following administration of the chemotherapeutic drug vincristine include increased mitoses, abnormal nuclear configurations, and fragmented nuclei due to binding of tubulin and inhibition of the mitotic spindle (Alleman and Harvey 1993). Iron deficiency, most commonly related to chronic blood loss, is associated with decreased heme synthesis resulting in an extra cell division and smaller than normal metarubicytes and erythrocytes (microcytosis). The cytoplasm of hemoglobin-deficient metarubricytes appears shaggy with poorly defined margins (Rebar 1993). Lead toxicosis also interferes with heme synthesis resulting in iron accumulation in erythroid precursors, which can be detected using iron stains such as Perls’ Prussian blue. The lesion is referred to as sideroblastic change and the affected erythroid cells are called siderocytes/sideroblasts (Travlos 2006b).
3D volume segmentation and reconstruction. Supervised image classification and automated quantification of superparamagnetic iron oxide nanoparticles in histology slides for safety assessment
Published in Nanotoxicology, 2021
Anna Bogdanska, Oliviero L. Gobbo, Yuri Volkov, Adriele Prina-Mello
Following the SPION biodistribution and pharmacokinetic study (Gobbo et al. 2015), the organs were fixed in 4% formalin and paraffin embedded. Samples from two animals per time point were used in this study. Four consecutive sections (5 µm thick) were collected at three levels. First section was stained with hematoxylin and eosin (H&E) stain for general toxicity studies (data not shown). The following two sections were stained with Perls’ Prussian blue (PPB) staining as detailed in previous publication with minor modifications (Edge et al. 2016a). Briefly, rehydrated sections were covered with equal parts of hydrochloric acid (HCL) and potassium ferrocyanide (K4[Fe(CN)6]·3H2O) for 20 min. Slides were washed in distilled water and counterstained with eosin for 5 min. Following incubation periods, slides were rinsed with distilled water before being dehydrated in a graded series of alcohol rinses. Slides were then cleared in two baths of xylene, sealed under coverslips with DPX mounting medium and examined under light microscope. To evaluate if any iron-positive staining is masked by the counterstain, third section was stained with PPB stain omitting eosin step in the outlined protocol (data not shown).
Butea monosperma (Lam.) Taub. Bark fractions protect against free radicals and induce apoptosis in MCF-7 breast cancer cells via cell-cycle arrest and ROS-mediated pathway
Published in Drug and Chemical Toxicology, 2020
Varinder Kaur, Manish Kumar, Ajay Kumar, Satwinderjeet Kaur
This assay was performed in order to examine the reduction of ferricyanide complex (Fe3+) into its ferrous form (Fe2+) by the extract/fractions, using the protocol described by Oyaizu (1986). In brief, various concentrations of the extract/fractions (25–400 µg/mL) along with 2.5 mL phosphate buffer and 2.5 mL potassium ferricyanide were mixed together in a glass test tube and incubated at 50 °C for 20 min; the assay was performed in triplicate. Following the incubation, 2.5 mL trichloroacetic acid (TCA) was added to the mixture in order to stop the reaction. The supernatant (2.5 mL) obtained after centrifugation at 3000 rpm for 10 min was mixed with 2.5 mL distilled water, followed by the addition of 0.5 mL ferric chloride (FeCl3) solution. The appearance of Perls’ Prussian blue coloration was spectrophotometrically recorded at 700 nm against the blank solution. An increase in the absorbance of the reaction mixture was considered an indicator of the reducing ability of the sample. The results of the assay were calculated using the formula given below:
Intracranial lymphatic drainage discharges iron from the ventricles and reduce the occurrence of chronic hydrocephalus after intraventricular hemorrhage in rats
Published in International Journal of Neuroscience, 2020
Wang Bocheng, Liang Chaofeng, Chen Chuan, He Haiyong, Huang Tengchao, Gao Qun, Guo Ying
The whole study was divided into three parts (Table 1 and Figure 2). In part I, 4 rats received 200 μl autologous blood intraventricular injection and 1 rats received saline injection as control. DCLNs were collected for Perls Prussian blue reaction at 6, 12, 24 and 48 h after injection. In part II, rats were divided into three groups. IVH were induced in 40 rats and 20 of them accepted DCLNs excision before the injection. The other six rats were control. The ventricles volume of survival rats were examined by MR at 28 days after injection. In part III, 24 rats accepted IVH and 12 of them suffered DCLNs excision before injection. Saline were injected into ventricles in nine rats as control. The ferritin level of surrounding ventricles were evaluated by western blotting at 3, 7 and 28 days after IVH.