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Meiotic Abnormalities in Infertile Males
Published in Carlos Simón, Carmen Rubio, Handbook of Genetic Diagnostic Technologies in Reproductive Medicine, 2022
Mireia Solé, Francesca Vidal, Joan Blanco, Zaida Sarrate
In contrast, studies of meiotic chromosomes using uniform staining have been implemented in some laboratories. Briefly, this protocol consists of mechanical disaggregation of the testicular tissue in a hypotonic solution (0.075 M KCl), followed by cell fixation using methanol:acetic acid (3:1). The cell suspension obtained is dropped onto slides, and samples are stained with Leishman stain (20%) for evaluation with an optical microscope (Figure 5.3). This methodology allows the analysis of chromosome features at prophase I, metaphase I, and metaphase II, identifying meiotic abnormalities in these stages (Figure 5.4).
Evaluation of sperm
Published in David K. Gardner, Ariel Weissman, Colin M. Howles, Zeev Shoham, Textbook of Assisted Reproductive Techniques, 2017
Kaylen M. Silverberg, Tom Turner
While observing sperm in a counting chamber or on a slide, additional cell types may also be seen. These include endothelial cells from the urethra, epithelial cells from the skin, immature sperm cells, and white blood cells. The most common and significant of these cell types is referred to collectively as “round cells.” These include immature sperm cells and white blood cells. In order to distinguish between them, an aliquot of semen can be placed in a thin layer on a slide and air-dried. The cells are fixed to the slide and stained using a Wright-Giemsa or Bryan-Leishman stain. When viewed under 400x or 1000X power, cell types may be differentiated primarily by their nuclear morphology. Immature sperm have one to three round nuclei within a common cytoplasm. Polymorphonuclear leukocytes may also be multinucleate, but the staining method will typically reveal characteristic nuclear bridges between their irregularly shaped nuclei (1). A peroxidase stain may be used to identify granulocytes and to differentiate them from the immature sperm. The presence of greater than 1 million white blood cells per one milliliter of semen may indicate an infection in the urethra or accessory glands, which provide the majority of the seminal plasma. Such infections could contribute to infertility (1, 11). As such, these samples must be cultured so that the offending organism can be identified and appropriate treatment can be instituted. Besides bacteria, white blood cells on their own can contribute to infertility. They can especially be a detrimental factor in the in vitro fertilization (IVF) process. Even though the white blood cells can be removed by centrifugation of the semen sample through a layer of silica beads, the toxins produced by the cells, called leukokines, may pass through the layer and concentrate in the medium below containing the sperm. If this sperm is to be used in the insemination of oocytes, the concentrated toxins will be in contact with the oocytes for several hours.
Detection of abnormal lymphocytes in the peripheral blood of COVID-19 cancer patients: diagnostic and prognostic possibility
Published in Hematology, 2022
Lobna Refaat, Mona S. Abdellateif, Ahmed Bayoumi, Medhat Khafagy, Eman Z. Kandeel, Hend A. Nooh
Peripheral blood (PB) samples were obtained from all participating subjects at admission during the routine workup of the patients. The blood samples were collected in Ethylenediaminetetraacetic acid (EDTA) vacutainers and prepared for undergoing complete blood count (CBC) analysis. Also, differential total leukocyte count (TLC) was done using SYSMEX XN1000 and SYSMEX XT 1800 analyzers, which included an absolute count of lymphocytes, monocytes, and neutrophils, eosinophils, basophils, and immature granulocytes. The immature granulocytes represented an automated count of promyelocytes, myelocytes, and metamyelocytes in the peripheral blood. Peripheral blood (PB) smears were done by spreading one drop of the blood on a slide and stained with Leishman stain. The WBC morphology was analyzed as changes from normal expected/baseline morphology. The blood films were examined by two experienced hematopathologists using Lecia light microscope with 100× oil-immersed magnifications.
Relationship between sputum periostin level and inflammatory asthma phenotypes in Egyptian patients
Published in Journal of Asthma, 2021
Maged Mohamed Refaat, Eman El Sayed, Wael Abd El-Fattah, Amr Helmy Elbanna, Hoda Mohamed El Sayed
Sputum analysis for inflammatory cells and periostin level. Sputum induction was done by nebulized hypertonic saline 3% and salbutamol for maximum 20 min. The sputum was isolated from saliva by forceps. One portion of the sputum was liquefied using ultrasonic waves then underwent ultracentrifugation. The supernatant was used for quantitative measurement of sputum level of periostin using ELISA technique by The Thermo Scientific™ Pierce™ Human Periostin ELISA Kit, Massachusetts, USA. The detection limit is 80 pg/ml. A phosphate-buffered saline solution was prepared with dithiothreitol (DTT), at 0.1% concentrations and was added to an equal volume of sputum to cause mucolysis. The mucus samples were stained with Leishman stain and total and differential cell counts were evaluated. Inflammatory phenotypes of asthma were then defined as eosinophilic if (sputum eosinophils ≥3% and sputum neutrophils <61%), neutrophilic if (sputum neutrophils ≥61% and sputum eosinophils <3%), paucigranulocytic if (sputum neutrophils <61% and sputum eosinophils <3%), or mixed granulocytic if (sputum neutrophils ≥61% and sputum eosinophils ≥3%; 19,20).
Apoptotic lymphocytes on peripheral smear and positional parameter values (VCS data) can suggest a diagnosis of infectious mononucleosis
Published in Infectious Diseases, 2019
Jayashree D. Kulkarni, Sanjay A. Pai
PS were stained with Leishman stain. The percentage of apoptotic lymphocytes, reactive lymphocytes and normal lymphocytes were obtained by doing a differential count of 100 consecutive lymphocytes. The VCS parameters, i.e. the mean and standard deviation (SD) of VCS of the lymphocytes and monocytes were reviewed. Because dengue is common, we wondered whether similar features of apoptosis and VCS data could be seen in dengue as well. Hence, we compared our data with 20 patients diagnosed with dengue fever and 20 healthy individuals.