Seminar 2 Notes
Seminar 2 Notes
Seminar 2 Notes
Ebersole
Timetable Seminar 2
Subject: Hematology ( 1 and 2) – ERYTHROCYTES
WEEK 1 Erythropoiesis, Hemoglobin, NO QUIZ YET
Hematocrit
WEEK 2 Erythropoiesis, Hemoglobin, Hematocrit –
50pts
READ YOUR PREVIOUS NOTES ALSO
ESR, RBC Indices, Morphology
and Inclusion Bodies
WEEK 3 Possible Continuation
Erythropoiesis
Grading of Polychromasia
Slight 1+
2+ 3+
4+
Radioisotopic Evaluation
Iron(59) Long term Radiation, Very high energy that do not permit good image
Iron(52) Excellent for imaging, tandem with cyclotron
Tc(99m) Most used, Low radiation, must be linked with sulfur, for Medullary Hematopoiesis
Cr(51) or Random Method for Red Cell Survival
In (111)
HEMATOPOIESIS
Red-iron bearing protein contained within the erythrocytes in normal blood for
transportation of oxygen and carbon dioxide and regulation of acid base balance
Hemoglobin Structure: with ____ heme molecules and ___ different pairs of globin
Heme Structure
With Carbon, Hydrogen and Nitrogen( Protoporphyrin IX) with central divalent
ferrous iron which reversibly combines with one oxygen
Globin Chains
Chromosome __ Chromosome __
___ amino acids ___ amino acids
Alpha(α), Zeta(δ) Beta(β) Gamma A(γa), Gamma G(γg),
Delta(δ), Epsilon(ε)
Heme Biosynthesis
Hemoglobin Synthesis
To efficiently bind oxygen in the lungs with high oxygen affinity and to unload
oxygen in the tissues with low oxygen affinity
Approximately ______ of oxygen is bound by ________ of hemoglobin
Relationship of oxygen is described in oxygen dissociation curve which is
_______ in shape
Low oxygen tension leading to low affinity for oxygen = TISSUES
o Shift to the left = higher oxygen affinity than normal
Lower body temperature
Multiple transfusion with low 2,3 DPG
Alkalosis ( High pH)
High Affinity to Oxygen Hemoglobin ( HbF, COHb, Hb Chesapeake)
o Shift to the right = lower oxygen affinity than normal
Increased body temperature
Increased 2,3 DPG ( Hypoxia, Anemia, High Altitude)
Acidosis ( Low pH)
Low Affinity to Oxygen hemoglobin ( Hb Kansas)
Bohr effect = A shift that is due to ___ ( Hydrogen concentration)
Haldane effect = Oxygenation of blood in the lungs displaces ___________ from
hemoglobin which increases the removal of carbon dioxide.
Forms of Hemoglobin
3. Chemical method (iron content method): 1gm of hgb is equal to 3.47mg of iron
Colorimetric methods
A. Visual
A1. Direct matching = tallquist, dare ( Acid hematin and Alkali hematin)
Acid hematin Alkali hematin
Ex. Sahli’s Hellige, Sahli-Adams, Osgood Ex. Clegg and King
Newcomer, Haldane, Haden-Hausser
B.Photoelectric
B1. Oxyhemoglobin method: ______________ for measuring plasma hemoglobin. It uses
20ul blood + n/150 ammonia solution / 0.1% Na2CO3.
B2. Cyanmethomoglobin or HiCN method: ___________________. It uses Drabkin’s
reagent + 20ul blood, read at 540nm. Sodium lauryl sulphate may be used to produce SLS-
methemoglobin which is non toxic
Principle:
Advantages:
1. Can measure almost all forms of Hgb except __________
2. Sample can be directly compared with HiCN standard
Disadvantages:
1. Affected by lipemia due to turbidity: corrected by adding 0.01mL of plasma to 5mL of
cyanmethemoglobin reagent and used it as patient blank
2. May affect leucocytosis ( >20,000) or thrombocytosis (> 700,000): corrected by the
reagent-sample solution can be centrifuged and the supernatant measured.
3. May affect by hemoglobins such as HbS and HbC which are resistant. This can be
corrected by making a 1:2 dilution with distilled water (1 part diluted sample plus 1 part
water) and multiplying the results from the standard curve by 2.
4. Also affected by globulinemia such as multiple myeloma and waldenstrom
macroglobulinemia due to increase in protein. To remedy, add 0.1 g of potassium
carbonate to the cyanmethemoglobin reagent. This is now modified by adding potassium
hydrogen phosphate in the reagent.
5. Cyanmethemoglobin reagent is sensitive to light
6. Carboxyhemoglobin takes 1 hour to be converted to HiCN
A. Macromethod
Method Anticoagulant
Wintrobe method Double oxalate
Haden’s modification method 1.1% sodium oxalate
Van Allen’s method 1.6% sodium oxalate
Sanford-Magath method 1.3% sodium oxalate
Bray’s method Heparin
Factors to be considered
1. Trapped plasma causes ______________ in cases of sickle cell anemia, macrocytic
anemias, hypochromic anemias, spherocytosis, and thalassemia by 1% to 3%.
2. Not recommended after blood loss due to false decrease
3. Dehydration and hemoconcentration causes false increased
4. Hemolysis and interstitial fluid false decreased
QUIZ 1 up to this
READ PREVIOUS NOTES ALSO associated with the topics assigned
Intrinsic factors
Plasma factors Increased albumin
Increased lecithin
Defibrination
Red cell factors Microcytosis
Spherocytosis
Polycythemia
Increased sickle cells and poikilocytes
Extrinsic factors
A. Temperature below 200c E. Long standing of blood
If two cell population is present, RDW is increased, Decreased RDW has little
significance, NV: 11.5% to 14.5%
120 days
survival rate
Same size of
a small
lymphocyte
11
Lecture Notes in Seminar 1 Chester F. Ebersole
Sideroblastic
Anemia
Red Cell Hemoglobin Content: due to the shape of the cell and to degenerative changes
or abnormalities of cell formation, particularly HEMOGLOBIN SYNTHESIS
Central Pallor
is about 1/3
Anemia of
Chronic
disease
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Lecture Notes in Seminar 1 Chester F. Ebersole
Dual Dimorphic
Population/ anemia
Heterogenous
population Post
Transfusion
Degree of Hypochromia
Hypochromasia Grading
+1 ½ of central pallor
+2 2/3 of the central pallor
+3 ¾ central pallor
+4 Thin rim of hemoglobin
Due to:
A. Lowered hemoglobin conc. B. Abnormal thinness
B. Morphologic
Megaloblastic anemia
Porphyria
Lead Poisoning
Iron Deficiency Anemia
Thalassemia
Sideroblastic Anemia
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Lecture Notes in Seminar 1 Chester F. Ebersole
Mcleod
Phenomenon
Vitamin E
deficiency
Hypothyroidism
__________/ Fragments of Extreme Disseminated
keratocyte/ RBCs varying fragmentation intravascular
Pyknocyte/ in size and produced by coagulation (DIC)
damage of
Helmet Cell shape and
RBCs by fibrin,
often display altered vessel Micro
pointed walls and angiopathic
extremities prosthetic hemolytic anemia
heart valves Physical Agents
Burns
Uremia
Renal Graft
Rejection
_________ RBC’s with Due to Disorders with
(Degmacyte) semicircular Splenic RBC inclusions
portion Pitting of
removed from Heinz
the cell Bodies
margin
*** Under
Schistocyte
Blister Cell Same as Bite Cell
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Lecture Notes in Seminar 1 Chester F. Ebersole
May be
curved or S
shape
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Lecture Notes in Seminar 1 Chester F. Ebersole
Megaloblastic
Anemia
Morphology Grade As
Polychromatophilia 1+ = 1 to 5 / field
Helmet Cell 2+ = 6 to 10 / field
Teardrop RBC 3+ = > 10/field
Acanthocyte
Schistocyte
Spherocyte
Poikilocytosis 1+ = 3 to 10 / field
Ovalocytes 2+ = 11 to 20/ field
Burr Cells 3+ = > 20 / fields
Bizarre Shaped RBC
Elliptocytes
Stomatocyte
Rouleaux 1+ = aggregates of 3 to 4 RBCs
2+ = aggregates of 5 to 10 RBCs
3+ = numerous aggregate with few RBCs
Sickle Cell Report as positive only
Basophillic Stippling
Pappenheimer bodies
Howell Jolly Bodies
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Lecture Notes in Seminar 1 Chester F. Ebersole
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Lecture Notes in Seminar 1 Chester F. Ebersole
QUIZ 2 Up to Here
READ PREVIOUS NOTES ASSOCIATED WITH THE ASSIGNED TOPICS
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Lecture Notes in Seminar 1 Chester F. Ebersole
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