Professional Documents
Culture Documents
By
Felicia Magee Tardy, M.S.,
MT (ASCP)
Department of Clinical Pathology
University of Mississippi Medical Center
Jackson, Mississippi
Introduction
Hematology: the study of blood and
blood forming tissues.
Blood consists of 55% plasma and
45% formed elements.
Formed elements include
erythrocytes, leukocytes, and
thrombocytes.
Erythrocytes
Normal range 4.25.5 million per mm3
in adults.
Biconcave shape.
Diameter 7
microns.
Cells for transport
of O2 and CO2.
Life span 120 days.
Leukocytes
Normal range 4 - 11
thousand per mm3
in adults.
Five types.
Size 8-20 microns.
Involved in fighting
infection,
combatting allergic
reactions, and
immune
responses.
Thrombocytes
Smallest cells in
the blood.
Normal range
130,000-400,000.
Active role in
coagulation and
hemostasis.
Routine Hematology
Anticoagulant of choice: EDTA
Automated Counting
Coulter Principle
Electrical impedance: resistance or
change in current when cell passes
between two electrodes in NaCl
solution.
Automated Counting
Flow Cytometry
Uses lasers to measure both forward
and side scatter.
Forward scatter measures size.
Side scatter measures granularity.
Sources of Error
Inadequate mixing of specimen.
Hemolyzed specimens.
Lipemic specimens.
Cold agglutinins.
Clotted specimens.
Platelet clumps or platelet
satellitosis.
Diluted specimens.
PLT
MPV
SEGS
LYMPHS
MONOCYTES
EOSINOPHILS
BASOPHILS
Erythrocytic
Maturation Series
Rubriblast
Prorubricyte
Rubricyte
Metarubricyte
Reticulocyte
Erythrocyte
RBC
Morphology
Elliptocytes
Target Cells
Tear Drops
Stomatocytes
Sickle Cells
Schistocytes
RBC
Inclusions
Howell-Jolly Bodies
Round, purple
inclusions in
RBCs.
Composed of DNA.
Commonly seen in
in patients with
hypofunctioning
spleens.
Splenectomy.
Basophilic Stippling
Numerous, small
purple inclusions
in RBCs.
Aggregates of
ribosomal RNA.
Most commonly
seen in lead
poisoning.
Pappenheimer Bodies
Clusters of dark
blue granules,
irregular in size
and shape.
Composed of iron
and ribosomal
RNA.
Seen in
sideroblastic and
hemolytic anemias.
Classifications of Anemias
Microcytic, Hypochromic
Iron deficiency
Sideroblastic
Chronic disease, Inflammation
Lead poisoning
Thalassemia trait
Microcytic, Hypochromic
Many RBCs smaller
than
nucleus of normal
lymphocytes,
increased central
pallor.
Iron deficiency,
thalassemias,
anemia of chronic
disease.
Classifications of Anemias
Normochromic
Hereditary Spherocytosis
Hereditary Elliptocytosis
PNH
G6PD deficiency
Aplastic anemia
Acute blood loss
Classifications of Anemias
Macrocytic
Vitamin B12 deficiency
Folate deficiency
Liver disease
Macrocytic RBCs
Most RBCs larger
than nucleus of
normal
lymphocytes,
increased MCV.
Folate or Vitamin
B12 deficiencies,
alcoholism, and
liver disease.
Reticulocytes
Immature RBCs.
Contain residual
ribosomal RNA.
Reticulum stains
blue using a
supravital stain
(new methylene
blue).
Counted and
expressed as % of
total red cells.
Reticulocyte Count
Uses supravital stain which stains
cells in the living state.
Retic % =
Hemoglobinopathies
Beta Chain Substitutions
Hgb S: Valine for glutamic acid
(6th position, beta chain)
Hgb C: Lysine for glutamic acid
(6th position, beta chain)
Hemoglobinopathies
Alkaline Electrophoresis
C
A2
S F A +
D
E
O
WBC
Evaluation
Myelocytic
Maturation Series
Myeloblast
Promyelocyte
Myelocyte
Metamyelocyte
Band Neutrophil
Segmented Neutrophil
Toxic Granulation
Increased
basophilic granules
in neutrophils.
Seen in severe
infections, burns,
malignancies, and
pregnancy.
Distinguish from
basophils.
Dohle Bodies
Sky blue inclusions
in cytoplasm of
neutrophils.
Seen in infections,
burns,
myleproliferative
disorders, and
pregnancy.
Composed of RER
and glycogen
granules.
Pelger-Huet Anomaly
Special Stains
PAS: ALL, Erytrholeukemia
LAP: CML v. Leukomoid Reaction
Myeloperoxidase: AML, AMMoL (weak+)
Sudan Black: AML, AMMoL (weak+)
Specific Esterase: AML, AMMoL
Non-specific Esterase: AMML, AMoL
TRAP: Hairy Cell Leukemia
TDT: ALL
Prussian Blue: Sideroblastic Anemia
CML versus
Leukomoid Reaction
Characteristic
LAP score
Toxic gran.
Dohle bodies
Philadelphia
chromosome
CML
Decreased
Decreased
Absent
May be present
Leukomoid
Increased
Increased
Present
Absent
WBC Disorders
Alder-Reilly: Large azurophilic granules,
increased mucopolysaccharides
Chediak-Higashi: Membrane defect of
lysosomes, large primary granules in
segs
May-Hegglin: Familial disorder with large
platelets and Dohle bodies
Pelger-Huet: Hyposegmented neutrophils
FAB Classifications
M1:
M2:
M3:
M4:
M5a:
M5b:
M6:
M7:
Chronic Myelogenous
Leukemia
FAB Classifications
L1: Small, uniform lymphoblasts
L2: Large, pleomorhphic
lymphoblasts
L3: Burkitts type (vacuolated and
deeply basophilic cytoplasm)
Acute Lymphoblastic
Leukemia
Chronic Lymphocytic
Leukemia
Body Fluids
Physical examination
RBC and WBC counts
WBC differential
Crystal examination*
Ropes test*
*Refers to synovial fluids only.
Quick Review
Differentiate between traumatic tap
and intracranial hemorrhage.
Differentiate between transudate and
exudate.
Crystal identification and associated
conditions.
Traumatic Tap v.
Intracranial Hemorrhage
Traumatic Tap
Intracranial Hemorrhage
Decreasing amounts
of blood with each
tube drawn
May contain clots
No xanthochromia
Supernatant clear
Transudates v. Exudates
Characteristic
Transudate
Exudate
Appearance
Clear, colorless
Yellow, turbid,
purulent, bloody
Specific gravity
<1.015
>1.015
Protein
<3 g/dL
>3 g/dL
LD
<200 IU
>200 IU
Cell count
<1000/uL
>1000/uL
Conditions Congestive
Infections,
Heart failure
Malignancies
Crystal Identification
Monosodium urate (uric acid)
Yellow when parallel to the compensator,
and blue when perpendicular.
Seen in gout.
Calcium pyrophosphate
Blue when parallel to the compensator,
and yellow when perpendicular.
Seen in pseudogout.
Crystal Identification
Good Luck!!!!