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Zagami

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    10
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    Italy

About Zagami

  • Birthday 01/05/1954

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  • AIM
    francesco_zagami@virgilio.it
  • MSN
    +393291809146

Profile Information

  • Gender
    Male
  • Biography
    https://independent.academia.edu/ZagamiFrancesco/CurriculumVitae
  • Location
    Tabaka Camillian's Mission Hospital
  • Occupation
    Clinical Pathologist
  • Real Name
    Zagami Francesco

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  1. Dear kackieanne, The coloring system you mentioned and used in Central America is called Diff-Quick and is a quick variant of the Romanosky coloring. This is an excellent stain for peripheral blood smears, but less sensitive for staining of malarial plasmodium. On the other hand, Field's coloring, as told by davierm, is more valid, of which I'm sending you the specifications for local production, which I proposed in my mission in Kinshasa (DRC) the Malaria Field Stain: Eosin stain: dissolve 1.3 g eosin Y by active stirring in 500 ml dH2O containing 12.6 g Na2HPO4.12H2O and 6.25 g KH2PO4 (Sorensen’s buffer at pH 6.8). Stand it at room temperature for 24 hours in a closed container; Polychrome methylene blue: dissolve 1.3 g methylene blue (C.I. 52015) in 500 ml dH2O containing 12.6 g of Na2HPO4.12H2O and boiled to dry powder to polychrome the dye is obtained. Dissolve the powder in 500 ml of dH2O containing 6.25 g KH2PO4.
  2. Not performing the microscopic examination anymore is a huge clinical damage because the cytology of the sediment is a fundamental in urological diagnostics, especially if performed on colored sediments. Even today I also prepare PAD's like the one for proteins (Albumin) described below: Protein urine strip (Protein-PAD) Reagent solution A: dissolve in 500 g dH2O, 130 g sodium citrate, 46.6 g citric acid, 0.8 g lauroyl sarcosine, and add 1 litre of methanol Reagent solution B: dissolve in 200 ml methanol 59.4 g magnesium sulfate, 0.5 g tetrabromophenolphthalein ethyl ester and fill up to 1 litre with methanol. Filter paper support: filter paper Whatman #1 is dipped in separate reagents solution and then each time is dried. Procedure: when the test strip obtained were dipped for 5 sec. into protein-containing urins, then uniformly green and blue colorations of the test zone were obtained, whereas when dipped, in same time, in protein-free urine gave a negative reaction indicated by a pale yellow color
  3. Posted August 28, 2016 The easy solution at the question, reported by colleenbennett, on the usefulness of cover the blood films with glass coverslips, not is ultimate because blood smears are preserve better without using coverslip. For storage stain peripheral blood smear slide a non-aqueous/permanent mounting medium (Entellan, DPX, Canada Balsam, and Neo-Mount) is recommended. As soon as the slide is completely dried, a few drops of a non-aqueous/permanent mounting medium are brought up on the preparation. Avoid air bubbles under the cover slip when the slide is covered. After drying time of 20 - 30 min, the smear can be examined under the microscope and stored in an archive. The so preserved preparation remains color stable for a minimum of 10 years. I have realized a permanent slides adding a drop of Canada balsam on the slide and adding a coverslip. Warm the slide to help the Canada balsam spread. Store flat and allow to harden over several days. We used for stain thin smear peripheral blood a quick procedure in which after fixing step, 1 min. in fixative solution (dissolve 0.002 g di Fast green in 1000 ml of absolute methanol or in denatured alcohol), the slide is rinse in tap water and dipped for 45 sec. in eosin G buffered (dissolve 0.95 g Yellow Eosin in phosphate buffer 0.2 M at pH 6.6 [dissolve in 300 ml warmed dH2O, 12.52 g Na2HPO4·2H2O, and 12.5 g di KH2PO4, check pH at 6.6, and dissolve 1 g NaN3 and make to 1 liter with dH2O]), then rinse the slide in tap water and dipped for 15 sec. in methylene blue-azure A buffered (dissolve 1.6 g methylene blue and 1.0 g azure A in phosphate buffer 0.2 M at pH 6.6 [dissolve in 300 ml warmed dH2O, 12.52 g Na2HPO4o2H2O, and 12.5 g di KH2PO4, check pH at 6.6, and make to 1 liter with dH2O]). Rinse the slide in tap water and allows to stay in vertical position for air dry.
  4. The Coding and Payment Guide for the Physical Therapist (CPT) is a numeric code that codify for specific analyte determined by means a specified methodology with a automated or manual procedure that for Bilirubin Total & Direct is 82247 for Bilirubin total (Neonatal Bilirubin) and 82248 for Bilirubin direct when measured with Jendrassik-Grof modified methodology applied on automated instrument. Through Bilirubin total reagent are measured in single determination conjugated, unconjugated and delta bilirubin (neonatal samples contain little or no direct δ-bilirubin) that produce a single result Bilirubin total (Neonatal Bilirubin) identified with 82247 code in CPT, therefore your approach is correct. I for measure neonatal bilirubin employ plasma obtained by capillary whole blood and as method a Jendrassik-Grof modified methodology two liquid stable reagents (Sulfanilic Acid Reagent : dissolve 5.58 g [32.2 mM] of sulfanilic acid to 15.5 -16 ml of HCl conc. 37% fuming and dilute to 500 ml con dH2O. Mix 250 ml [3.5 M] of DMSO with 250 ml [4.44 M] of ethylene glycol and fill up to 500 with dH2O. Mix the DMSO/ethylene glycol solution with the sulfanilic acid solution and store in polyethylene opaque at 4 °C stable 1 year. The solvent mixture of dimethyl sulfoxide, and ethylene glycol besides as solvent for the total bilirubin assay, eliminates interference from hemolysis [neonatal plasma] up to 10 g/L of hemoglobin; Sodium nitrite solution pH 8.0 : dissolve in 60 ml dH2O, 0.752 g [109 mM] of sodium nitrite, and 0.240 g [16.9 mM] of disodium hydrogen phosphate anhydrous [Na2HPO4], then 0.010 – 0.040 g [0.96 mM] ethylenediaminetetraacetic acid tetrasodium salt dihydrate, and fill up to 100 ml with dH2O with final pH about 8.0. Store under refrigeration, at 4 °C, in brown glass-stoppered bottle, resulting stable for 1 year, but must be discard if it becomes tinged with yellow. In aqueous solution the sodium nitrite is immediately degrade and convert itself to nitric oxide, while in a pH neutral or slightly alkaline solution, the sodium nitrite is stable). Ratio reagents SAR:sample:SNS is 1:0.1:0.01 and read Abs at 555 nm
  5. The Sysmex XN-Series multiparameter automated hematology analyzer, consist of a new channel named the WDF channel. This channel can differentiate leukocytes from cells treated with specific reagents containing detergents and fluorescent stains, by using the 2-parameter flowcytometric method. The scattergrams of the 2 channels have different patterns due to the differences in the reagents used as well as differences in the hardware and software. In particular, the WDF channel differentiates between lymphocytes and monocytes and enhances the separation capacity.
  6. Welcome to the forums Zagami :)

    1. Zagami

      Zagami

      Dear Admin Cliff,

      I have never changed my email address for an opportunity in scientific communication, which would be

      compromised if changed.

      Therefore, it seems strange how your previous email communication was returned as undeliverable.

      Sorry for the inconvenience

      Best Regards

      Francesco Zagami MD

       
       

       

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