The Wright-Giemsa stain - Secrets revealed
ABSTRACT The colorful story of the development of the Wright-Giemsa stain is retold. Dramas are replayed, secrets are exposed, and laurels are properly returned to scientists long forgotten. The delicately balanced chemical composition of the stain, once enigmatic, is defined. Finally, an attempt is made to unshroud some of the mystery surrounding the staining method; tried-and-true procedures and useful snippets of laboratory lore are provided. Scientific explanations and reliable methods aside, however, the stain continues to possess a certain mystique, seemingly consisting of equal parts technical expertise, art, and magic.
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ABSTRACT: Monocyte/macrophage adhesion to biomaterials, correlated with foreign body response, occurs through protein-mediated surface interactions. Albumin-selective perfluorocarbon (FC) biomaterials are generally poorly cell-conducive because of insufficient receptor-mediated surface interactions, but macrophages bind to albumin-coated substrates and also preferentially to highly hydrophobic fluorinated surfaces. Bone marrow macrophages (BMMO) and IC-21, RAW 264.7, and J774A.1 monocyte/macrophage cells were cultured on FC surfaces. Protein deposition onto two distinct FC surfaces from complex and single-component solutions was tracked using fluorescence and time-of-flight secondary ion mass spectrometry (ToF-SIMS) methods. Cell adhesion and growth on protein pretreated substrates were compared by light microscopy. Flow cytometry and integrin-directed antibody receptor blocking were used to assess integrins critical for monocyte/macrophage adhesion in vitro. Albumin predominantly adsorbs onto both FC surfaces from 10% serum. In cultures preadsorbed with albumin or serum-dilutions, BMMO responded similar to IC-21 at early time points. Compared with Teflon AF, plasma-polymerized FC was less permissive to extended cell proliferation. The beta(2) integrins play major roles in macrophage adhesion to FC surfaces: antibody blocking significantly disrupted cell adhesion. Albumin-mediated cell adhesion mechanisms to FC surfaces could not be clarified. Primary BMMO and secondary IC-21 macrophages behave similarly on FC surfaces, regardless of preadsorbed protein biasing, with respect to adhesion, cell morphology, motility, and proliferation.Journal of Biomedical Materials Research Part A 02/2009; 88(2):503-19. DOI:10.1002/jbm.a.31886 · 2.83 Impact Factor
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ABSTRACT: We report the novel, heterozygous AE1 mutation R730C associated with dominant, overhydrated, cation leak stomatocytosis and well-compensated anemia. Parallel elevations of red blood cell cation leak and ouabain-sensitive Na(+) efflux (pump activity) were apparently unaccompanied by increased erythroid cation channel-like activity, and defined ouabain-insensitive Na(+) efflux pathways of nystatin-treated cells were reduced. Epitope-tagged AE1 R730C at the Xenopus laevis oocyte surface exhibited severely reduced Cl(-) transport insensitive to rescue by glycophorin A (GPA) coexpression or by methanethiosulfonate (MTS) treatment. AE1 mutant R730K preserved Cl(-) transport activity, but R730 substitution with I, E, or H inactivated Cl(-) transport. AE1 R730C expression substantially increased endogenous oocyte Na(+)-K(+)-ATPase-mediated (86)Rb(+) influx, but ouabain-insensitive flux was minimally increased and GPA-insensitive. The reduced AE1 R730C-mediated sulfate influx did not exhibit the wild-type pattern of stimulation by acidic extracellular pH (pH(o)) and, unexpectedly, was partially rescued by exposure to sodium 2-sulfonatoethyl methanethiosulfonate (MTSES) but not to 2-aminoethyl methanethiosulfonate hydrobromide (MTSEA) or 2-(trimethylammonium)ethyl methanethiosulfonate bromide (MTSET). AE1 R730E correspondingly exhibited acid pH(o)-stimulated sulfate uptake at rates exceeding those of wild-type AE1 and AE1 R730K, whereas mutants R730I and R730H were inactive and pH(o) insensitive. MTSES-treated oocytes expressing AE1 R730C and untreated oocytes expressing AE1 R730E also exhibited unprecedented stimulation of Cl(-) influx by acid pH(o). Thus recombinant cation-leak stomatocytosis mutant AE1 R730C exhibits severely reduced anion transport unaccompanied by increased Rb(+) and Li(+) influxes. Selective rescue of acid pH(o)-stimulated sulfate uptake and conferral of acid pH(o)-stimulated Cl(-) influx, by AE1 R730E and MTSES-treated R730C, define residue R730 as critical to selectivity and regulation of anion transport by AE1.AJP Cell Physiology 04/2011; 300(5):C1034-46. DOI:10.1152/ajpcell.00447.2010 · 3.67 Impact Factor
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ABSTRACT: With recent advances in molecular techniques, collecting blood from birds has become a common practice among field ornithologists. There are a variety of techniques for collecting blood samples and numerous caveats for how samples should be processed, depending on the research question being asked. Currently, few resources are available for individuals learning how to collect blood from birds or needing more information about how to process blood samples. Here, I describe commonly used methods for collecting, processing, and storing blood for particular research objectives, and provide answers to frequently asked questions about blood collection. The information provided is intended primarily for investigators working with passerines, but many techniques and suggestions are applicable to other avian taxa.RESUMENCon los recientes avances en las técnicas moleculares, la colecta de sangre de las aves se ha convertido en una práctica común entre los ornitólogos de campo. Hay una variedad de técnicas para colectar muestras de sangre y numerosas advertencias de cómo deberán ser procesadas, en función a la pregunta de investigación. En la actualidad, hay pocos recursos disponibles para personas que están aprendiendo como colectar sangre de las aves, o para los que necesitan más información acerca de cómo procesar las muestras de sangre. Aquí, describo los métodos comúnmente utilizados para colectar, procesar y almacenar la sangre, dependiendo de los objetivos de la investigación en particular, y proveo respuestas a las preguntas más frecuentes acerca de la colecta de sangre. La información proporcionada está dirigida principalmente a los investigadores que trabajan con aves paseriformes, pero muchas técnicas y sugerencias son aplicables a otros grupos taxonómicos de aves.Journal of Field Ornithology 12/2011; 82(4). DOI:10.1111/j.1557-9263.2011.00338.x · 1.20 Impact Factor