Podcast
Questions and Answers
What is the primary purpose of hemoglobin electrophoresis?
What is the primary purpose of hemoglobin electrophoresis?
Which of the following factors could lead to a falsely elevated hemoglobin measurement?
Which of the following factors could lead to a falsely elevated hemoglobin measurement?
What correction should be made if a sample contains both Hb S and Hb C?
What correction should be made if a sample contains both Hb S and Hb C?
Which hemoglobin type is considered the slowest during cellulose acetate electrophoresis?
Which hemoglobin type is considered the slowest during cellulose acetate electrophoresis?
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What pH range is considered optimal for the primary screening procedure using cellulose acetate?
What pH range is considered optimal for the primary screening procedure using cellulose acetate?
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What is the effect of placing hemoglobin in an alkaline medium?
What is the effect of placing hemoglobin in an alkaline medium?
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Which method is mentioned as an alternative to hemoglobin electrophoresis?
Which method is mentioned as an alternative to hemoglobin electrophoresis?
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What is the significance of hemoglobin being sensitive to light?
What is the significance of hemoglobin being sensitive to light?
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Study Notes
Hemoglobin Errors
- High WBC count >20 x 109/L
- High platelet count >700 x 109/L
- Lipemia (presence of large amount of lipid)
- Cells containing Hb S and Hb C
- Abnormal globulins (found in plasma cell myeloma or Waldenstrom macroglobulinemia) precipitate in reagent
Correction Procedures
- Centrifuge reagent sample solution, then measure the supernatant
- Add 0.01 mL of the patient's plasma to 5mL of cyanmethemoglobin reagent, using this solution as a reagent blank
- Make a 1:2 dilution with distilled water (1 part diluted sample +1 part water) and multiply results by 2 from the standard curve
- Add 0.1 Gram of potassium carbonate to the cyanmethemoglobin reagent. Commercially available cyanmethemoglobin reagent is modified with KH2PO4 salt.
Reminders
- Cyanmethemoglobin reagent is sensitive to light, store in brown bottle or dark place.
- Another technique using sodium lauryl sulfate (SLS) transforms hemoglobin to SLS-methemoglobin; this method does not produce toxic wastes.
- HemoCue is a commercial handheld system to measure hemoglobin concentration
- Hemoglobin is converted to azidemethemoglobin in photometry, reading levels at 570 nm and 880 nm
Hemoglobin Electrophoresis
- Electrophoresis is the movement of charged particles in an electric field
- Cellulose acetate (pH 8.4-8.6) is the primary screening procedure for detecting variants in hemoglobin.
- Hemoglobin is a negatively charged molecule in an alkaline buffer (8.4-8.6).
- During electrophoresis, Hemoglobin molecules travel to the anode (+) due to their negative charge.
- Fastest (normal Hb) = HbA1 / HbA
- Note that fast hemoglobins= Hg that travels further than A1, HgF, HgB, etc..
- Slowest = HgA2, C, E, O Arab, Charlem
- Note Hb S, Hb D, and Hb G migrate to the same area in cellulose acetate electrophoresis.
- If hemoglobin is placed in an alkaline medium, it will assume a negative charge and travel to the anode (+).
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Description
This quiz covers the identification of hemoglobin errors, including abnormal blood counts and the presence of specific hemoglobin types. It also details correction procedures using cyanmethemoglobin reagent and important reminders for handling these reagents safely.
