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Pneumocystis carinii
Published in Peter D. Walzer, Robert M. Genta, Parasitic Infections in the Compromised Host, 2020
Peter D. Walzer, C. Kurtis Kim, Melanie T. Cushion
The second type of P. carinii stains stain trophozoites, intracystic bodies, and all intermediate forms. Giemsa stain is the prototype, but similar results can be obtained with Wright-Giemsa and polychrome methylene blue (305,310, 311). We favor Diff Quik, a variant of the Wright-Giemsa stain, because it is very rapid (can be accomplished in less than 1 min) and gives staining results similar to those achieved with the standard Giemsa stain (137,261). Recent studies indicate that P. carinii can also be seen with Wright's and Gram's stains in lung imprint smears and bronchoalveolar lavage fluid (311-316), but it is unclear what specific role these stains will have in diagnosis of this infection.
Pathology—Patient with possible Hirschsprung disease: Case study
Published in Victoria A. Lane, Richard J. Wood, Carlos A. Reck-Burneo, Marc A. Levitt, Pediatric Colorectal and Pelvic Surgery, 2017
Victoria A. Lane, Richard J. Wood, Carlos A. Reck-Burneo, Marc A. Levitt
The Diff-Quik stain consists of three solutions: Diff-Quik fixative reagent: triarylmethane dye, methanolDiff-Quik solution I (eosinophilic): xanthene dye, pH buffer, sodium azideDiff-Quik solution II (basophilic): thiazide dye, pH buffer
A Diagnostic Approach to Pneumocystis jiroveci Pneumonia
Published in Johan A. Maertens, Kieren A. Marr, Diagnosis of Fungal Infections, 2007
Abigail Orenstein, Henry Masur
Indirect Stains. Although trophozoite forms can occasionally be stained with the Gram-Weigert and PAS techniques, true “indirect” stains are primarily taken up by non-cyst forms, i.e., intracystic bodies (sporozoites) and (extracystic) trophozoites. Cysts can be visualized by their inability to take up stain and therefore appear “negatively” stained. Indirect stains can be performed more rapidly than the direct stains, but require more time and experience to interpret more information, although adequate skill can be achieved without extensive training. Care must be taken so as to not confuse organisms with host cells and debris; neutrophils entrapped in mucus can appear to be clusters of organisms. However, if correctly identified, the presence of neutrophils can be helpful in assessing prognosis. Wright-Giemsa, the prototypical indirect stain, can be completed within 30 minutes. A modified version, Diff-Quik, is inexpensive and takes only several minutes to prepare. Polychrome methylene blue is another indirect stain, although not commonly used.
Quercetin improves the imbalance of Th1/Th2 cells and Treg/Th17 cells to attenuate allergic rhinitis
Published in Autoimmunity, 2023
Xia Ke, Ziqi Chen, Xiaoqiang Wang, Houyong Kang, Suling Hong
Quercetin (purity ≥ 95%, Figure 1A), Ovalbumin (OVA), aluminum hydroxide, phorbol myristate acetate (PMA), ionomycin, and Brefeldin A (BFA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Diff-Quik Stain solution was purchased from Sbjbio (Nanjing, China). Hematoxylin and eosin staining solutions were purchased from Phygene (Fuzhou, China). Periodic acid-Schiff (PAS) staining kit was purchased from Beyotime (Shanghai, China). Glemsa stain working solution was purchased from Leagene (Beijing, China). All ELISA kits were purchased from FineTest (Wuhan, China). TRIzol reagent was purchased from Invitrogen (Carlsbad, CA, USA). PrimeScript RT Master Mix and Realtime PCR Master Mix (SYBR Green) were purchased from KeyGEN biotech (Nanjing, China). CD4+ T cell isolation kit (mouse) was purchased from NovoBiotechnology (Beijing, China). The fixation/permeabilization concentrate was purchased from eBioscience (San Diego, CA, USA). Antibodies used in flow cytometry were all purchased from Biolegend (San Diego, CA, USA). Antibodies used in western blot were all purchased from Abcam (Cambridge, MA, USA). BCA kit and ECL reagent were purchased from Beyotime (Shanghai, China).
PreservCyt Is an Optimal Fixative that Permits Cytologic and Molecular Analyses of Vitreoretinal Lymphoma Biopsies
Published in Ocular Immunology and Inflammation, 2021
Mona Meng Wang, Wei Jian Tan, Tong Seng Lim, Anita Sook Yee Chan
Pfeiffer cells pre-fixed in different fixatives (PreservCyt, HOPE, PAXgene, Shandon, Allprotect or RNAlater) and three vitreous clinical patient samples pre-fixed in PreservCyt were sent for cytological analysis at the Singapore General Hospital (SGH), Department of Anatomical Pathology and Cytology. Vitreous samples were centrifuged using Thermo Scientific Shandon Cytospin (Shandon Scientific Ltd, Cheshire, UK)9 and cytospin slides were obtained for Diff Quik (DQ, Baxter, McGraw Park, IL) and Papanicolaou (PAP) stains using standard diagnostic10 and previously published IHC protocols.11,12 In brief, single staining for the B-cell markers CD20 and CD79a, the T-cell marker CD3, and the proliferation index marker Ki-67 was performed using a Bond Polymer Refine Detection Kit (Leica Biosystems) and diaminobenzidine staining. Pfeiffer cells and clinical cytospin slides were visualised under a BX42 Olympus microscope (60
Acute pulmonary and splenic response in an in vivo model of whole-body low-dose X-radiation exposure
Published in International Journal of Radiation Biology, 2019
Stephanie Puukila, Stacy Muise, James McEvoy, Tara Bouchier, Antony M. Hooker, Douglas R. Boreham, Neelam Khaper, Dani-Louise Dixon
For splenocyte isolation, tissue was homogenized in PBS and 0.8% M ammonium chloride was used to lyse erythrocytes. Samples were centrifuged at 540 g and cell pellets were resuspended in RPMI media with 25 mM HEPES and NaHCO3 (Sigma Aldrich, USA, cat#R5886), 2 mM L-alanyl-glutamine (Sigma Aldrich, USA), and 100 U Penstrep (Thermo Fisher Scientific, USA). Cells were seeded in two 96-well plates and one 6 well plate at 1 × 106 cells per well. In control wells cells were grown in media, while in the remaining wells, 40 μg/mL 2× lipopolysaccharide (LPS; Sigma Aldrich, USA) or 4 μg/mL 2× concanavalin A (conA; Sigma Aldrich, USA) was used to stimulate proliferation at 37 °C and 5% CO2. Following incubation for 1 or 48 hours, 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) reagent (Promega, USA) was added to the 96-well plates and absorbance measured at 490 nm after 1 hour. MTS is a colorimetric method for quantification of viable cells in proliferation. Following 48 hour incubation of the 6-well plate, cells were collected, fixed in 4% paraformaldehyde and smeared on Superfrost Plus slides. Slides were stained with Diff-Quik (Thermo Fisher Scientific, USA) for cell differential analysis.