Determination of Hb values

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Determination of Hb values

• M. Ss. Program
• CLS 541
• Lab-3-

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Determination of Hb values

• Haemoglobin can be measured by manual or
  automatic.
• Two method are common use
• 1- haemiglobincyanid.
• (HicNCyanmethaemoglobin )
• 2-Oxyhaemoglobin (Hbo2method).

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• A major a advantage of the HicN method is
  availability of stable and reliable reference
  preparation.
• Other methods Sahlis acid haematin and
  alkalin-haematin method.

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Cyanmethaemoglobin method

• Use Drabkins solution which contain potassium
  cyanide and potassium ferricynide at PH7-7.4.
• Hb oxidation to methaemoglbin by potassium
  ferricynide then combine to potassium cyanide to
  form cyanmethaemoglobin which measure at 540 nm
  by spectrophotometer.

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Normal rage of Hb in g/dl

• Hb g/dl (Ax64500xD. F)(44XDx1000x10)
• Men 15 2 g/dl
• Women 13.5 1.5g/dl
• At birth 18 4g/dl

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R.B.C, W.B.C and Platelet count

• The cell blood count can used manual or
  automatic.
• A variety of automated instruments for
  performing blood counts have been developed and
  are now widespread use.
• Manual cell counts using counting chambers it is
  not recommended for routine because will be too
  few to ensure a precise.

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• Improved Neubauer counting chamber contain 9
  1mmx1mm ruled areas each contain a volume of
  0.1µl of diluted blood examine at 40x objective.
• For W.B.C count use the acidic acid with violet
  stain as the diluents but for R.B.C and platelets
  use the formal citrate as the diluents .

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• Normal rage for R.B.C count
• For men 5 0.5 x10 12/L.
• For women 4.3 0.5 x10 12/L.
• Normal range of W.B.C count
• For men and women 7.0 3.0X109L.
• Normal range of platelets count
• For men and wmen 150-400 x 109L.

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Packed Cell Volume (PCV) determination and red
cell indices

• Two methods
• 1-macro-method of wintrobe which is no longer in
  routine use.
• 2-micro-haemotocrite which the centrifuge used
  for capillary tubes provides a centrifugal force
  of c12000g and 5min.

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• PCV for men 0.45 0.05 L/L.
• PCV for women 0.41 0.05 L/L.
• PCV for infant 0.60 0.15 L/L.
• MCV (PCVX10) R.B.C.
• MCV92 9 FL.
• MCH (Hbg/dlx10) R.B.C.
• MCH 29.5 2.5 Pg.
• MCHC (Hb PCV) X 100.
• MCHC 33 1.5 0R g/dL.

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ESR DETERMINATION

• The recommended method is Westergren method.
• The test is performed on venous blood.
• It conventional to set up sedimentation rate
  tests at room temperature(18-25C).
• The usual practice is to collect the blood
  dirctly into the citrate solution.
• The test can be carried out equally well with
  blood anticoagulated with EDTA within24h.

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• If the specimem is kept at 4C,provided that
  1volume of 109mmol/l(23g/l) trisodium citrate is
  add to 4volumes of blood immediately befor the
  test is performed.
• the blood will draw it up into the Westergren
  tube to 200mm mark, place the tbe exactly
  vertical and leave undistrubed for exactly
  60min.the result expressed as ESRX min in 1h.
• ESR for menlt7010-14(gt70c30).
• ESR for womenlt7012-20(gt70c35).

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Reticulocyte count

• Reticulocytes are immature red cells.
• They contine remnants of ribosomal ribonucleic
  acid.
• To stain the reticulocytes use basic dyes such as
  azure B,brilliant creslyle blue or new
  methylene blue to form a blue or purple
  precipitate of granules of filments (unfixed
  prepartion).
• reticulocytes can be divided into four groups
  according maturation (group I until groupIV).

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• The group I which contain a large clump of
  reticun but group IV with few granules of
  reticulin group.
• The ripening process in thought to take 2-3 days.
• The number of reticulocytes in peripheral blood
  is a fairly accurate reflection of erythropoetic
  activity.
• Normal 0.05 - 2.5.
• Infant 2 - 5.