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Red cell countRed cell count
MethodsMethods
A. Manual diluting method.
Principle:
Using a diluting fluid that destruct all blood cells
leaving only RB...
ReagentsReagents
Gower’s solution: Sod. Sulphate,
Glacial acetic acid and D.W
Hayem’s solution: mercuric
chloride, NaCl,...
EquipmentsEquipments::
Hemocytometer
Cover slip
Pipettes
ProceduresProcedures::
1. Dilution:
Add 2 ml of hayem’s solution to clean
and dry tube.
Add 10 µl of the sample (whole ...
2. Charging
Charge the hemocytometer between the
cover slip and the counting chamber.
Leave the cells to settle down.
...
16 Tertiary
squares
Reading (countingReading (counting((
????? Cell /square.
CalculationCalculation::
Erythrocytes / µl of blood= cells counted per
square х 10 (depth) х 5 х200(dilution)
OR
Erythro...
B. Other automatic methodsB. Other automatic methods::
Electronic cell counter
Flow cell cytometers : depends on laser
l...
RBCS MORPHOLOGYRBCS MORPHOLOGY
AND ABNORMALITIESAND ABNORMALITIES
RBCs morphologyRBCs morphology::
Biconcave disc
Flexibility, due to actin and
spectrin
Mamalian RBCs are non
nucleated.
Po...
Poultry RBCs are elliptical nucleated.
Fish RBCs are round nucleated.
Dog Cat Equine Bovine Sheep Goat
Size 7µm 5.8 µm 5.7 µm 5.5 µm 4.5 µm < 4 µm
Central palor V. clear V.
Slight
Lack SLight
...
Abnormalities in RBCs morphology:Abnormalities in RBCs morphology:
 Abnormalities in size (Anisocytosis):
variation in si...
Variation of sizeVariation of size::
Polychromatophilic red
blood cell , macrocyte
Pseudomacrocytosis
(reticulocyte)
True macrocytosis
Causes???
Pernicious ane...
AnisocytosisAnisocytosis
 Abnormalities in aggregation:
Agglutination: RBCs aggregated in the form of Clumps or clusters
( not in chains)
Causes :...
Abnormalities in RBCs shape:Abnormalities in RBCs shape:
(Pokilocytosis)(Pokilocytosis)
Acanthocyte ( spur cellAcanthocyt...
Creanation – Echinocyte – burr cell:
(blunt to sharp projections, regularly distributed, even in
size and diameter)
Cause...
AcanthocyteEcchinocyte
Target cell ( codocytes) or leptocytes (folded
cell):
( centeral hemoglobinized area separated from pereferal
hemoglobini...
Tear drop cell (Dacrocytes):
pear shaped with elongated blunt or pointed projection.
Causes:- Is thought to be due to dis...
Tear drop cell (Dacrocytes):Tear drop cell (Dacrocytes):
Sickle cell ( Drepanocyte):
Elongated, Fusiform, Spindle shaped or Crescent , and lack
central palor.
Due to polymerlizat...
Spherocytes: Small darkly stained RBC
that loss central pallor
Incidence:- 1- Characteristic in IMHA
(Due to incomplete p...
 Stomatocytes : - Red cells in which the central pallor
is straight or appears as a curved rod-shaped slit.
 The cells a...
Abnormalities in Hb contentAbnormalities in Hb content::
Hypochromacia
Polychromacia.
Hypochromacia
RBCs inclusions
( structures in or on RBCs):
Basophilic stippling.
Heinz bodies.
Howell- jolly bodies.
Parasites, babe...
RBCs inclusionsRBCs inclusions::
Basophilic stippling.
Invivo aggregation of ribosoms
into small basophilic
granules
Inci...
Howell- jolly bodies:- nuclear reminants in
erythrocytes ( small, round, dark blue inclusions of
variable Size
Shape
Site...
Heinz bodies:Heinz bodies: Appear asAppear as small, eccentric, pale refractile
structure within RBC membrane and commonly...
While by vital stains ( NMB, brillent cresyl blue)
Appear as blue structures.
Babesia in RBCsBabesia in RBCs
Babesia in RBCs using Wright’s stain.
Nucleated RBCs ( Erythroblastosis)Nucleated RBCs ( Erythroblastosis)
associated with regenerative anemia, spleenic dysfunc...
RBC indices include:
Mean Corpuscular Volume (MCV(
Mean Corpuscular Hemoglobin (MCH(
 Mean Corpuscular Hemoglobin Conce...
MCVMCV
Mean cell volume
MCV is average size of RBC
MCV = Hct x 10
RBC (millions)
If 60-77 fL, normal range(canine), RB...
MCHMCH
MCH is average weight of hemoglobin
per RBC.
MCH = Hgb x 10
 RBC (millions)
Unite is pg
Normal canine range is...
MCHCMCHC
MCHC is average hemoglobin
concentration per RBC
MCHC = Hgb x 100
Hct (%)
If MCHC is normal, cell described as...
RDWRDW
Most automated instruments now
provide an RBC Distribution Width
(RDW)
An index of RBC size variation
May be use...
Section 3 rb cs
Section 3 rb cs
Section 3 rb cs
Section 3 rb cs
Section 3 rb cs
Section 3 rb cs
Section 3 rb cs
Section 3 rb cs
Section 3 rb cs
Section 3 rb cs
Section 3 rb cs
Section 3 rb cs
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Section 3 rb cs

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Transcript of "Section 3 rb cs"

  1. 1. Red cell countRed cell count
  2. 2. MethodsMethods A. Manual diluting method. Principle: Using a diluting fluid that destruct all blood cells leaving only RBCs intact. With the aid of a hemocytometer, counts the number of cells expressed in millions per micro liter of blood.
  3. 3. ReagentsReagents Gower’s solution: Sod. Sulphate, Glacial acetic acid and D.W Hayem’s solution: mercuric chloride, NaCl, sod. Sulphate and D.W.  Physiological saline (0.85%)
  4. 4. EquipmentsEquipments:: Hemocytometer Cover slip Pipettes
  5. 5. ProceduresProcedures:: 1. Dilution: Add 2 ml of hayem’s solution to clean and dry tube. Add 10 µl of the sample (whole anti- coagulated blood) and mix. Dilution factor: 1:200
  6. 6. 2. Charging Charge the hemocytometer between the cover slip and the counting chamber. Leave the cells to settle down. The depth factor is 110 3. Counting: Count the RBCs in 5 medium squares (4 at the 4 corners & 1 in the center) inside the central large square. Count those cells in L-shape manner
  7. 7. 16 Tertiary squares
  8. 8. Reading (countingReading (counting(( ????? Cell /square.
  9. 9. CalculationCalculation:: Erythrocytes / µl of blood= cells counted per square х 10 (depth) х 5 х200(dilution) OR Erythrocytes / µl of blood= cells counted per square х 10 (depth)х 25х200 (dilution) = …………………. Cells / µl or cells / mm3
  10. 10. B. Other automatic methodsB. Other automatic methods:: Electronic cell counter Flow cell cytometers : depends on laser lights.
  11. 11. RBCS MORPHOLOGYRBCS MORPHOLOGY AND ABNORMALITIESAND ABNORMALITIES
  12. 12. RBCs morphologyRBCs morphology:: Biconcave disc Flexibility, due to actin and spectrin Mamalian RBCs are non nucleated. Poultry RBCs are elliptical nucleated. Fish RBCs are round nucleated.
  13. 13. Poultry RBCs are elliptical nucleated. Fish RBCs are round nucleated.
  14. 14. Dog Cat Equine Bovine Sheep Goat Size 7µm 5.8 µm 5.7 µm 5.5 µm 4.5 µm < 4 µm Central palor V. clear V. Slight Lack SLight Rouleux mild degree Marked Common Rare in health and disease condition Anisocytosis Uniform size Mild Common Characteristics Pokilocytosis Slight Characteristics
  15. 15. Abnormalities in RBCs morphology:Abnormalities in RBCs morphology:  Abnormalities in size (Anisocytosis): variation in size of cells due to presence of micro &/or Macrocytes among normal cells. Macrocytosis (↑MCV) : 1- True macrocytosis (vit. B12 and folate deficiency) erythrocyte maturing factors 2-Pseudo macrocytosis (regenerative response of bone marrow in case of blood loss.) Microcytosis (↓MCV): iron deficiency or failure in absorption or utilization of iron ( deficiency of Cu, Vit C, vit. B6
  16. 16. Variation of sizeVariation of size::
  17. 17. Polychromatophilic red blood cell , macrocyte Pseudomacrocytosis (reticulocyte) True macrocytosis Causes??? Pernicious anemia
  18. 18. AnisocytosisAnisocytosis
  19. 19.  Abnormalities in aggregation: Agglutination: RBCs aggregated in the form of Clumps or clusters ( not in chains) Causes :- Autoimmune mediated diseases). Staphylococcal infection ( Protein A) Rouleaux: RBCs arranged in the form of chain, like stock of coins. Normal in horse and to a Mild degree in dog and increase in Tumors, infection and inflammation (hyperfibrinogenemia) Rare in ruminant either in health or disease conditions How can they differentiated
  20. 20. Abnormalities in RBCs shape:Abnormalities in RBCs shape: (Pokilocytosis)(Pokilocytosis) Acanthocyte ( spur cellAcanthocyte ( spur cell)) (Irregular spine, uneven in size, numbers, diameter and distribution) Causes:- Irregular projections , formed when erythrocyte membrane contain excess cholesterol compared to phospholipids, Incidence:- common in severe liver disease.
  21. 21. Creanation – Echinocyte – burr cell: (blunt to sharp projections, regularly distributed, even in size and diameter) Causes and incidence: in case of renal insufficiency, uremia, artifact in lab. Techniques(due to slow air drying)
  22. 22. AcanthocyteEcchinocyte
  23. 23. Target cell ( codocytes) or leptocytes (folded cell): ( centeral hemoglobinized area separated from pereferal hemoglobinized area by clear zone) due to increased surface Area : volume ratio membrane folding and target cell formation Causes:- Occurs from low Hb content as in case of iron deficiency anemia, liver disease, regenerative Anemia. Seen in dog with increasersd serum cholesterol Level.
  24. 24. Tear drop cell (Dacrocytes): pear shaped with elongated blunt or pointed projection. Causes:- Is thought to be due to distortion of RBCs as they travel through vasculature of abnormal B.M or spleen. Incidence:-  myelofibrosis, metastatic tumor in the bone marrow, extramedullary hematopoiesis, dyserythropoiesis, megaloblastic anemias,
  25. 25. Tear drop cell (Dacrocytes):Tear drop cell (Dacrocytes):
  26. 26. Sickle cell ( Drepanocyte): Elongated, Fusiform, Spindle shaped or Crescent , and lack central palor. Due to polymerlization of Hb Causes:- Occurs in Hemoglobinopathies (mutation of hemoglobin gene).
  27. 27. Spherocytes: Small darkly stained RBC that loss central pallor Incidence:- 1- Characteristic in IMHA (Due to incomplete phagocytosis) 2- Hereditary spherocytosis. Schistocyte or Keratocyte: are red cell fragments which are formed when fibrin strands come in contact with circulating red cells. The strands cut a small piece from the original cell. Incidence:-Several microangiopathic diseases, including disseminated intravascular coagulation, intravascular truma and vascular neoplasm (hemangiosarcoma)
  28. 28.  Stomatocytes : - Red cells in which the central pallor is straight or appears as a curved rod-shaped slit.  The cells appear as "smiling face" or fish mouth. Incidence:- occurs in hereditary stomatocytosis, liver disease and may be as artifact in thick blood film.
  29. 29. Abnormalities in Hb contentAbnormalities in Hb content:: Hypochromacia Polychromacia. Hypochromacia
  30. 30. RBCs inclusions ( structures in or on RBCs): Basophilic stippling. Heinz bodies. Howell- jolly bodies. Parasites, babesia and anaplasma.  Nucleated RBCs.
  31. 31. RBCs inclusionsRBCs inclusions:: Basophilic stippling. Invivo aggregation of ribosoms into small basophilic granules Incidence:- occurs in lead poisoning in Dog and responsive anemia in ruminant .
  32. 32. Howell- jolly bodies:- nuclear reminants in erythrocytes ( small, round, dark blue inclusions of variable Size Shape Site Incidence:-  spleenic dysfunction  Spleenectomy  regenerative anemia
  33. 33. Heinz bodies:Heinz bodies: Appear asAppear as small, eccentric, pale refractile structure within RBC membrane and commonly seem to protrude slightly from RBC marigin. ( with routine stain( It called refractile body in cat and reach 10 % normally . Causes:- oxidative denaturation of Hb.by oxidative drugs and compound ( Onion, garlic and Zn & Cu toxicity.  it increase susceptibility for Hemolysis.
  34. 34. While by vital stains ( NMB, brillent cresyl blue) Appear as blue structures.
  35. 35. Babesia in RBCsBabesia in RBCs Babesia in RBCs using Wright’s stain.
  36. 36. Nucleated RBCs ( Erythroblastosis)Nucleated RBCs ( Erythroblastosis) associated with regenerative anemia, spleenic dysfunction
  37. 37. RBC indices include: Mean Corpuscular Volume (MCV( Mean Corpuscular Hemoglobin (MCH(  Mean Corpuscular Hemoglobin Concentration (MCHC( RBC Distribution Width (RDW(
  38. 38. MCVMCV Mean cell volume MCV is average size of RBC MCV = Hct x 10 RBC (millions) If 60-77 fL, normal range(canine), RBCs considered normocytic If < 60 fL are microcytic If > 77 fL are macrocytic Not reliable when have marked anisocytosis
  39. 39. MCHMCH MCH is average weight of hemoglobin per RBC. MCH = Hgb x 10  RBC (millions) Unite is pg Normal canine range is 19-24 pg
  40. 40. MCHCMCHC MCHC is average hemoglobin concentration per RBC MCHC = Hgb x 100 Hct (%) If MCHC is normal, cell described as normochromic If MCHC is less than normal, cell described as hypochromic There are no hyperchromic RBCs Normal canine range is 32-36
  41. 41. RDWRDW Most automated instruments now provide an RBC Distribution Width (RDW) An index of RBC size variation May be used to quantitate the amount of anisocytosis on peripheral blood smear Increase in case of regenerative anemia
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