Peripheral blood smear examination
Dr Hemang Mendpara
DNB pediatrics
Choithram Hospital & Research Centre
Indore
www.dnbpe...
• Hemogram:
measured and
calculated
parameters
• Histograms:
size distribution of
WBC, RBC and Plt
• Cytogram: WBC
differe...
A well made peripheral smear is thick at one end and progressively thinner
at the opposite end. The "zone of morphology" (...
Slide fixation and staining
1. Romanowsky staining
Leishman's stain : a polychromatic stain
•Methanol : fixes cells to sli...
PBS examination requires a systematic approach in
order to gather all possible information.
In addition, all specimens mus...
• 1. Macroscopic view : quality of the smear
• 2.The microscopic analysis
• begins on lower power (10x),
• to assess cellu...
(a) Ten microscopic fields are examined in a vertical direction
from bottom to top or top to bottom
(b) slide is horizonta...
1. RBC
• Size
• Shape
• Color
• Arrangement
• Inclusions
• Young RBCs
2. WBC
• Total counts
• Differential counts
• I:T ra...
•A fairly accurate estimate of the WBC count (cells/mL)
can be obtained by counting the total number of
leukocytes in ten ...
Morphologic Evaluation of Red Blood Cells
Biconcave disc
Diameter : 7 ~ 8 μm
Central pallor occupy 1/3 rd of total
Size : ...
Microcytic hypochromic red cells
Decreased size and Hb
content (MCH) and conc
(MCHC). Expanded
central zone of pallor
Iron...
Iron deficiency anemia
www.dnbpediatrics.com
Thalassemia trait
www.dnbpediatrics.com
Megaloblastic anemia (PS)
Macrocyte
Large RBCs
• size > 8.5 mm,
• MCV > 95 fL
• Normal MCH
•Normal newborn
•Chromosomal di...
Elliptocytes or ovalocytes
Ovalocytes are due to abnormal membrane
cytoskeleton found in hereditary elliptocytoisis
www.dn...
seen when there is extramedullary erythropoiesis
Tear drop cells / dacrocytes
• Osteopetrosis
• Myelofibrosis
• Bone marro...
Polychromasia
Blue-gray coloration of RBCS.
Due RNA remnants
Increased - Increased erythropoietic
activity. Decreased - Hy...
Sickle cell anemia
Irregular, curved cells
with pointed ends
Hb S hemoglobinopathies (sickle cell anemia, Hb SC
disease, H...
Spherocytosis
Hereditary spherocytosis
•ABO incompatibility
•Autoimmune hemolytic anemia (warm
antibody type)
•Infections ...
Acanthocytes or spur cells, are spherical cells with blunt-tipped
or club-shaped spicules of different lengths projecting ...
Echinocytes
"Sea urchin cells,
crenated cells, burr cells"
Post-splenectomy,
uremia, hepatitis of the newborn, malabsorpti...
Mechanical damage to RBCs from fibrin deposits
 DIC
 MicroAngiopathic HA
 TTP/HUS
 prosthetic heart valves
 severe va...
hemolyic anemias
Hallmark: Presence of schistocytes , fragmented RBC
www.dnbpediatrics.com
Uniconcave RBC,
slitlike area of central
pallor
Hereditary or acquired hemolysis.
Hereditary stomatocytosis, alcoholic
cir...
HA due to red cell enzyme defects – bite or blister cells
• Glucose 6 phosphate dehydrogenase
(G-6-PD) deficiency
• Unstab...
Target cell
Peripheral rim of pallor surrounding central hyperchromia
Target cells are commonly seen in
•Hemoglobin C
•Sic...
Irregular RBC
agglutination/
clumping
Anti-RBC antibody, paraprotein. Cold
agglutinin disease, autoimmune hemolytic
anemia...
Roulex formation
Seen in case of high level of fibrinogen, immunoglobulins,
intra venous administration of plasma volume e...
• multiple blue-purple inclusions attached to the inner surface of the red cell membrane.
visible in supravitally stained ...
Small (1 mm), round,
dense, basophilic
bodies in RBCs.
Splenectomized patients,
Functional asplenia,
Anatomical absence of...
Basophillic strippling
• Lead poisoning
• Iron deficiency anemia
• Thalassemia
Are abnormal aggregrates of ribosome and po...
• Smaller then Howell jolly body
• Stain with Prussian blue stain
• Suggest iron over load
www.dnbpediatrics.com
WBC Morphology
www.dnbpediatrics.com
www.dnbpediatrics.com
Manual differential counts
• These counts are done in the same area as
WBC and platelet estimates with the red cells
barel...
•If 10 or more nucleated RBC's (NRBC) are seen,
correct the
• White Count using this formula:
•Corrected WBC Count =
WBC x...
• Left-shift: non-segmented neutrophil > 5%
– Increased bands Means acute infection, usually
bacterial
www.dnbpediatrics.c...
• Basophils are increased in the blood in
– Myeloproliferative disorders (e.g., chronic myelogenous leukemia)
– Hypersensi...
• Morphologically abnormal eosinophils are seen in
– Myelodysplastic syndrome
– Megaloblastic anemias
• Eosinophils are in...
Band cells
www.dnbpediatrics.com
Leukemic myeloblast
www.dnbpediatrics.com
Leukemic myeloblast stained with peroxidase
Note the AUER ROD
www.dnbpediatrics.com
Burkitt lymphoma
www.dnbpediatrics.com
Large, coarse, dark purple, azurophilic granules that occur in
the cytoplasm of most granulocytes. These are
characteristi...
Chédiak-Higashi granules are very large red or blue
granules that appear in the cytoplasm of granulocytes,
lymphocytes, or...
Variably sized (0.1 to 2.0 um) and shaped, blue or grayish-
blue cytoplasmic inclusions usually found near the
periphery o...
May-Hegglin anomaly
Neutrophils contain small basophilic cytoplasmic granules
which represent aggregated ribosomes. Leukop...
Neutrophilic toxic granulation
Small dark blue to purple granules resembling primary
granules in the cytoplasm of metamyel...
Platelets
Neubars chamber : count platelets in 64 small
squares
Counts * 250 = total platelets
Normal counts 4.5 to 5.5 la...
www.dnbpediatrics.com
:Giant plateletsPlatelet morphology
www.dnbpediatrics.com
Platelet satellitism
www.dnbpediatrics.com
Macrocytosis with giant platelets (MDS,
5q- syndrome)
www.dnbpediatrics.com
Disadvantages of the Peripheral Blood Smear
Provides information that cannot be obtained from automated
cell counting. How...
Merozoits
www.dnbpediatrics.com
Schizonts are commonly seen in P. vivax infection and appear as large
bodies containing 12 to 24 nuclei and a loose pigmen...
Schuffer’s dots seen in plasmodium vivex
www.dnbpediatrics.com
Cresent shaped gametocyte charectaristiclly seen in p.falciparum
malaria
www.dnbpediatrics.com
www.dnbpediatrics.com
www.dnbpediatrics.com
Eucheria bancrofti
www.dnbpediatrics.com
www.dnbpediatrics.com
www.dnbpediatrics.com
OSMOTIC FRAGILITY
TEST
• Defination:
• it is a test that measures the resistance to
hemolysis of red blood cells (RBC) by
...
• Isotonic (physiological) solution – 0.9 %
NaCl
• RBC burst in hypotonic (< 0.9 % NaCl),
and shrink (crenate) in hyperton...
• NORMAL RANGE:
• - hemolysis onset at: 0.45-0.5 % NaCl
• - hemolysis complete at: 0.3-0.33 % NaCl
• FACTORS AFFECTING OSM...
- Hereditary spherocytosis
- Acquired spherocytosis
- Hemolytic anemia (HDN)
- Malaria
- Severe pyruvate kinase deficiency...
www.dnbpediatrics.com
• Thalassemia
• Sickle cell anemia (hemoglobinopathy)
• Iron deficiency anemia
• Asplenia
• Liver disease
www.dnbpediatric...
www.dnbpediatrics.com
www.dnbpediatrics.com
introduction
www.dnbpediatrics.com
Principle of test
• Deoxygenated Hb-S is insoluble in the presence of a
concentrated phosphate buffer solution and forms a...
Procedure
• 1. sodium diethanoid 200mg+10 ml distilled
water
• 2. sickling buffer solutions
• Take 2 part of 1st solution ...
Result
www.dnbpediatrics.com
www.dnbpediatrics.com
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  1. 1. Peripheral blood smear examination Dr Hemang Mendpara DNB pediatrics Choithram Hospital & Research Centre Indore www.dnbpediatrics.com
  2. 2. • Hemogram: measured and calculated parameters • Histograms: size distribution of WBC, RBC and Plt • Cytogram: WBC differential CBC on automated analyzers Flagging for abnormalities necessitates a manual PBS reviewwww.dnbpediatrics.com
  3. 3. A well made peripheral smear is thick at one end and progressively thinner at the opposite end. The "zone of morphology" (area of optimal thickness for light microscopic examination) should be at least 2 cm in length. The smear should occupy the central area of the slide and be margin-free at the edges www.dnbpediatrics.com
  4. 4. Slide fixation and staining 1. Romanowsky staining Leishman's stain : a polychromatic stain •Methanol : fixes cells to slide •methylene blue stains RNA,DNA blue-grey color •Eosin stains hemoglobin, eosin granules orange-red color •pH value of phosphate buffer is very important www.dnbpediatrics.com
  5. 5. PBS examination requires a systematic approach in order to gather all possible information. In addition, all specimens must be evaluated in the same manner, to assure that consistent information is obtained. www.dnbpediatrics.com
  6. 6. • 1. Macroscopic view : quality of the smear • 2.The microscopic analysis • begins on lower power (10x), • to assess cellular distribution, staining quality, and to select an area where the RBCs are barely touching each other. •On hi-dry (40x), to obtain a WBC estimate. All of the detailed analysis of the cellular elements using high power or oil immersion. PBS examination - preliminary www.dnbpediatrics.com
  7. 7. (a) Ten microscopic fields are examined in a vertical direction from bottom to top or top to bottom (b) slide is horizontally moved to the next field (c) Ten microscopic fields are counted vertically. (d) procedure is repeated until 100 WBCS have been counted (zig zag motion) Scanning technique for WBC differential count and morphologic evaluation www.dnbpediatrics.com
  8. 8. 1. RBC • Size • Shape • Color • Arrangement • Inclusions • Young RBCs 2. WBC • Total counts • Differential counts • I:T ratio • Abnormal WBC 3. Platelets • Counts • Abnormality 4. Parasites Evaluation of PBS www.dnbpediatrics.com
  9. 9. •A fairly accurate estimate of the WBC count (cells/mL) can be obtained by counting the total number of leukocytes in ten 40X microscopic fields, dividing the total by 10, and multiplying by 3000. These estimates should approximate that obtained by the cell analyzer. WBC estimation on peripheral smear www.dnbpediatrics.com
  10. 10. Morphologic Evaluation of Red Blood Cells Biconcave disc Diameter : 7 ~ 8 μm Central pallor occupy 1/3 rd of total Size : approx. same as nucleus of mature lymphocyte www.dnbpediatrics.com
  11. 11. Microcytic hypochromic red cells Decreased size and Hb content (MCH) and conc (MCHC). Expanded central zone of pallor Iron deficiency, thalasemia trait Anemia of chronic disease www.dnbpediatrics.com
  12. 12. Iron deficiency anemia www.dnbpediatrics.com
  13. 13. Thalassemia trait www.dnbpediatrics.com
  14. 14. Megaloblastic anemia (PS) Macrocyte Large RBCs • size > 8.5 mm, • MCV > 95 fL • Normal MCH •Normal newborn •Chromosomal disorders (e.g., Trisomy 21) •Drug associated anticonvulsants, antidepressants, sulpha, chemotherapeutic agents, estrogen and antiretroviral agents) •Dyserythropoiesis •Myelodysplasia •Preleukemia •Hypothyroidism •Liver disease •Folate deficiency •BI2 deficiency www.dnbpediatrics.com
  15. 15. Elliptocytes or ovalocytes Ovalocytes are due to abnormal membrane cytoskeleton found in hereditary elliptocytoisis www.dnbpediatrics.com
  16. 16. seen when there is extramedullary erythropoiesis Tear drop cells / dacrocytes • Osteopetrosis • Myelofibrosis • Bone marrow infiltrated with hematological or non-hematological malignancies • Iron deficiency anemia • Pernicious anemia www.dnbpediatrics.com
  17. 17. Polychromasia Blue-gray coloration of RBCS. Due RNA remnants Increased - Increased erythropoietic activity. Decreased - Hypoproliferative states. Hemolytic anemias •Blood loss anemias •Recovering anemia www.dnbpediatrics.com
  18. 18. Sickle cell anemia Irregular, curved cells with pointed ends Hb S hemoglobinopathies (sickle cell anemia, Hb SC disease, Hb S-beta-thalassemia, Hb SD disease, hb Memphis /S disease) * Don’t be confused with fragmented RBC www.dnbpediatrics.com
  19. 19. Spherocytosis Hereditary spherocytosis •ABO incompatibility •Autoimmune hemolytic anemia (warm antibody type) •Infections (e.g., EBV, CMV, E. coli, Sepsis/Urosepsis) •Severe burns •DIC and HUS www.dnbpediatrics.com
  20. 20. Acanthocytes or spur cells, are spherical cells with blunt-tipped or club-shaped spicules of different lengths projecting from their surface at irregular intervals. Acanthocytes Acanthocytes are seen in •Hereditary abetalipoproteinemia •Hereditary acanthocytosis •End stage liver disease •Anorexia nervosa •Malnutrition •Post splenectomy •Intravenous hyperalimentation particularly with intralipid infusion www.dnbpediatrics.com
  21. 21. Echinocytes "Sea urchin cells, crenated cells, burr cells" Post-splenectomy, uremia, hepatitis of the newborn, malabsorption states, after administration of heparin, pyruvate kinase def phosphoglycerate kinase deficiency, uremia, HUS. Crenated / Burr cells / Echinocytes (Echinocytes, or burr cells or crenated red cells, in contrast, have shorter, sharp to blunt spicules of uniform length which are more evenly spaced around their periphery). www.dnbpediatrics.com
  22. 22. Mechanical damage to RBCs from fibrin deposits  DIC  MicroAngiopathic HA  TTP/HUS  prosthetic heart valves  severe valvular stenosis  malignant hypertension  March hemoglobinuria  myelofibrosis  hypersplenism Schistocyte – fragmented RBC  normal newborns  bleeding peptic ulcer  Aplastic Anemia  pyruvate kinase def  Vasculitis  Glomerulonephritis  renal graft rejection  severe burns  iron deficiency, thalassemia www.dnbpediatrics.com
  23. 23. hemolyic anemias Hallmark: Presence of schistocytes , fragmented RBC www.dnbpediatrics.com
  24. 24. Uniconcave RBC, slitlike area of central pallor Hereditary or acquired hemolysis. Hereditary stomatocytosis, alcoholic cirrhosis, acute alcoholism, obstructive liver disease, malignancy, severe infection, treated acute leukemia, artifact. Stomatocyte – fish mouth cell www.dnbpediatrics.com
  25. 25. HA due to red cell enzyme defects – bite or blister cells • Glucose 6 phosphate dehydrogenase (G-6-PD) deficiency • Unstable hemoglobin variants • Congenital Heinz body anemia Suggest oxidative stress www.dnbpediatrics.com
  26. 26. Target cell Peripheral rim of pallor surrounding central hyperchromia Target cells are commonly seen in •Hemoglobin C •Sickle cell disease •Hemoglobin E •Hemoglobin H disease •Thalassemias •Iron deficiency anemia •Liver disease •Target cells are seen with most of the hemoglobinopathies www.dnbpediatrics.com
  27. 27. Irregular RBC agglutination/ clumping Anti-RBC antibody, paraprotein. Cold agglutinin disease, autoimmune hemolytic anemia, macroglobulinemia, hypergammaglobinemia RBC autoagglutination www.dnbpediatrics.com
  28. 28. Roulex formation Seen in case of high level of fibrinogen, immunoglobulins, intra venous administration of plasma volume expanders like dextran www.dnbpediatrics.com
  29. 29. • multiple blue-purple inclusions attached to the inner surface of the red cell membrane. visible in supravitally stained smears. • are precipitated normal or unstable hemoglobin usually secondary to oxidant stress. • G6PD deficiency • Unstable hemoglobinopathy • Cong. Bite cell anemia Heinz body www.dnbpediatrics.com
  30. 30. Small (1 mm), round, dense, basophilic bodies in RBCs. Splenectomized patients, Functional asplenia, Anatomical absence of spleen Howell Jolly bodies Howell-Jolly bodies are small round bodies composed of DNA, about 1 µm in diameter, usually single and in the periphery of a red cell. They are readily visible on the Wright- Giemsa-stained smear. The spleen is responsible for the removal of nuclear material in the red cells, so in absence of a functional spleen, nuclear material is removed ineffectively. Howell-Jolly bodies are seen in •Post splenectomy •Functional asplenia •Anatomical absence of spleen www.dnbpediatrics.com
  31. 31. Basophillic strippling • Lead poisoning • Iron deficiency anemia • Thalassemia Are abnormal aggregrates of ribosome and polyribosomes www.dnbpediatrics.com
  32. 32. • Smaller then Howell jolly body • Stain with Prussian blue stain • Suggest iron over load www.dnbpediatrics.com
  33. 33. WBC Morphology www.dnbpediatrics.com
  34. 34. www.dnbpediatrics.com
  35. 35. Manual differential counts • These counts are done in the same area as WBC and platelet estimates with the red cells barely touching. • This takes place under 100 (oil) using the zigzag method. • Count 100 WBCs including all cell lines from immature to mature. Reporting results • Absolute number of cells/µl = % of cell type in differential x white cell count www.dnbpediatrics.com
  36. 36. •If 10 or more nucleated RBC's (NRBC) are seen, correct the • White Count using this formula: •Corrected WBC Count = WBC x 100/( NRBC + 100) •Example : If WBC = 5000 and 10 NRBCs have been counted •Then 5,000 100/110 = 4545.50 •The corrected white count is 4545.50 www.dnbpediatrics.com
  37. 37. • Left-shift: non-segmented neutrophil > 5% – Increased bands Means acute infection, usually bacterial www.dnbpediatrics.com
  38. 38. • Basophils are increased in the blood in – Myeloproliferative disorders (e.g., chronic myelogenous leukemia) – Hypersensitivity reactions – Mastocytosis – Xeroderma pigmentosa – Hypothyroidism www.dnbpediatrics.com
  39. 39. • Morphologically abnormal eosinophils are seen in – Myelodysplastic syndrome – Megaloblastic anemias • Eosinophils are increased in the following conditions:  Allergies  Parasitic infestations  Infections  Acute leukemia  Myeloproliferative diseases  Hypereosinophilic syndrome  Drug-associated www.dnbpediatrics.com
  40. 40. Band cells www.dnbpediatrics.com
  41. 41. Leukemic myeloblast www.dnbpediatrics.com
  42. 42. Leukemic myeloblast stained with peroxidase Note the AUER ROD www.dnbpediatrics.com
  43. 43. Burkitt lymphoma www.dnbpediatrics.com
  44. 44. Large, coarse, dark purple, azurophilic granules that occur in the cytoplasm of most granulocytes. These are characteristically found in the Alder-Reilly anomaly and in patients with mucopolysaccharidoses Alder-Reilly anomaly www.dnbpediatrics.com
  45. 45. Chédiak-Higashi granules are very large red or blue granules that appear in the cytoplasm of granulocytes, lymphocytes, or monocytes in patients with the Chédiak- Steinbrinck-Higashi syndrome. It is a rare autosomal recessive disorder Chédiak-Higashi www.dnbpediatrics.com
  46. 46. Variably sized (0.1 to 2.0 um) and shaped, blue or grayish- blue cytoplasmic inclusions usually found near the periphery of the cell. Dohle bodies are lamellar aggregates of rough endoplasmic reticulum, which appear in the neutrophils, bands, and metamyelocytes of patients with infection, burns, uncomplicated pregnancy, toxic states, or during treatment with hematologic growth factors - G-CSF. Döhle bodies www.dnbpediatrics.com
  47. 47. May-Hegglin anomaly Neutrophils contain small basophilic cytoplasmic granules which represent aggregated ribosomes. Leukopenia and large platelets are also found. An autosomal dominant trait, the May-Hegglin anomaly is associated with a mild bleeding tendency, but not by an increased susceptibility to infection www.dnbpediatrics.com
  48. 48. Neutrophilic toxic granulation Small dark blue to purple granules resembling primary granules in the cytoplasm of metamyelocytes, bands, and segmented neutrophils during inflammatory states, burns, and trauma, and upon exposure to hematopoietic growth factors. It is usually accompanied by a shift to the left and vacuolations in the cytoplasm (toxic vacuolations) and Dohle bodies. www.dnbpediatrics.com
  49. 49. Platelets Neubars chamber : count platelets in 64 small squares Counts * 250 = total platelets Normal counts 4.5 to 5.5 lakh Common Causes of Thrombocytopenia •Decreased production −Aplastic anemia −Acute leukemia −Viral infections *Parvovirus *CMV −Amegakaryocytic thrombocytopenia (AMT) •Increased destruction −Immune thrombocytopenia *Idiopathic thrombocytopenic purpura (ITP) *Neonatal alloimmune thrombocytopenia (NAITP) −Disseminated intravascular coagulation (DIC) −Hypersplenism Thrombocytosis • Reactive thrombocytosis  Post infection  Inflammation  Juvenile rheumatoid arthritis  Collagen vasvular disease • Essential thrombocythemia www.dnbpediatrics.com
  50. 50. www.dnbpediatrics.com
  51. 51. :Giant plateletsPlatelet morphology www.dnbpediatrics.com
  52. 52. Platelet satellitism www.dnbpediatrics.com
  53. 53. Macrocytosis with giant platelets (MDS, 5q- syndrome) www.dnbpediatrics.com
  54. 54. Disadvantages of the Peripheral Blood Smear Provides information that cannot be obtained from automated cell counting. However, some limitations are: • Experience is required to make technically adequate smears. • There is a non-uniform distribution of white blood cells over the smear, with larger leukocytes concentrated near the edges and lymphocytes scattered throughout. • There is a non-uniform distribution of RBCs over the smear, with small crowded red blood cells at the thick edge and large flat red blood cells without central pallor at the feathered edge www.dnbpediatrics.com
  55. 55. Merozoits www.dnbpediatrics.com
  56. 56. Schizonts are commonly seen in P. vivax infection and appear as large bodies containing 12 to 24 nuclei and a loose pigmented body. This photograph shows an early schizont of P. vivax on the left and mature schizonts www.dnbpediatrics.com
  57. 57. Schuffer’s dots seen in plasmodium vivex www.dnbpediatrics.com
  58. 58. Cresent shaped gametocyte charectaristiclly seen in p.falciparum malaria www.dnbpediatrics.com
  59. 59. www.dnbpediatrics.com
  60. 60. www.dnbpediatrics.com
  61. 61. Eucheria bancrofti www.dnbpediatrics.com
  62. 62. www.dnbpediatrics.com
  63. 63. www.dnbpediatrics.com
  64. 64. OSMOTIC FRAGILITY TEST • Defination: • it is a test that measures the resistance to hemolysis of red blood cells (RBC) by osmotic stress created by hypotonic solutions • RBC are exposed to a series of saline (NaCl) solutions with increasing dilution • The sooner hemolysis occurs, the greater is osmotic fragility of RBC www.dnbpediatrics.com
  65. 65. • Isotonic (physiological) solution – 0.9 % NaCl • RBC burst in hypotonic (< 0.9 % NaCl), and shrink (crenate) in hypertonic solutions (> 0.9 % NaCl) • Red cells are suspended in a series of tubes containing hypotonic solutions from 0.9 to 0 % NaCl. Degree of hemolysis measured for each NaCl concentration. www.dnbpediatrics.com
  66. 66. • NORMAL RANGE: • - hemolysis onset at: 0.45-0.5 % NaCl • - hemolysis complete at: 0.3-0.33 % NaCl • FACTORS AFFECTING OSMOTIC FRAGILITY • - cell membrane permeability • - surface-to-volume ratio www.dnbpediatrics.com
  67. 67. - Hereditary spherocytosis - Acquired spherocytosis - Hemolytic anemia (HDN) - Malaria - Severe pyruvate kinase deficiency www.dnbpediatrics.com
  68. 68. www.dnbpediatrics.com
  69. 69. • Thalassemia • Sickle cell anemia (hemoglobinopathy) • Iron deficiency anemia • Asplenia • Liver disease www.dnbpediatrics.com
  70. 70. www.dnbpediatrics.com
  71. 71. www.dnbpediatrics.com
  72. 72. introduction www.dnbpediatrics.com
  73. 73. Principle of test • Deoxygenated Hb-S is insoluble in the presence of a concentrated phosphate buffer solution and forms a turbid suspension that can be easily visualized. • Normal Hemoglobin A and other hemoglobins remain in solution under these conditions. These different qualitative outcomes allow for the detection of sickle cell disease and its traits. • SICKLEDEX® uses Saponin to lyse the red blood cells. Sodium Hydrosulfite then reduces the released hemoglobin. Reduced Hb-S is insoluble in the concentrated phosphate buffer and forms a cloudy, turbid suspension. Thus give a positive result. www.dnbpediatrics.com
  74. 74. Procedure • 1. sodium diethanoid 200mg+10 ml distilled water • 2. sickling buffer solutions • Take 2 part of 1st solution and 3 part of 2nd solution • Have one drop of blood on slide and put single drop of mixed solution • Wait for 30 mins • Watch under microscopewww.dnbpediatrics.com
  75. 75. Result www.dnbpediatrics.com
  76. 76. www.dnbpediatrics.com

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