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About Zagami

  • Birthday 01/05/1954

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  • Location
    Tabaka Camillian's Mission Hospital
  • Occupation
    Clinical Pathologist
  • Real Name
    Zagami Francesco

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  1. 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.
  2. 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
  3. 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.
  4. Welcome to the forums Zagami :)


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