Unit 3.5 DIFFERENTIAL COUNT Flashcards
This involves both quantitative and qualitative study of the types of leukocytes.
DIFFERENTIAL COUNT
It also includes quantitative and qualitative study of thrombocytes as well as morphologic abnormalities of erythrocytes
DIFFERENTIAL COUNT
represents the percentage distribution of the different WBCs
DIFFERENTIAL COUNT
METHODS OF BLOOD SMEAR PREPARATION
Preferred for bone marrow preparations
Coverslip Method
It has the advantage of superior leukocyte distribution
Coverslip Method
NOT for routine peripheral blood smear preparation.
Coverslip Method
Coverslip Method Procedure:
Place (?) at the center of coverslip held between thumb & forefinger
Invert another coverslip rotated (?)
As soon as the blood spreads, pull apart the 2 coverslips in a (?).
1 drop of blood
45O
firm and steady motion
Glass Slide-Coverslip Method
Place 1 drop of blood near (?) of a glass slide. Place coverslip.
As soon as the blood spreads, (?) the coverslip along the surface of the slide
one end
push forward
Most convenient and most commonly used.
Two-Glass Slide Method (Wedge slide/Push smear)
Two-Glass Slide Method (Wedge slide/Push smear) Disadvantages:
1. Tendency of larger cells to settle at the slide edges & (?)
2. (?) of nucleated cells
3. (?) to cells
feathered ends
Poor distribution
Greater trauma
Utilizes the wedge procedure
Miniprep/Hemaprep
A precision blood spreader prepares dual smears simultaneously at a constant angle and speed.
Miniprep/Hemaprep
Glass slides are placed on a platform inside the instrument
Hemaspinner
Instrument spins causing the slide to be covered with a monolayer of cells beam of sensor light to detect blood distribution
Hemaspinner
For finding reactive/immature or abnormal cells that are present in small numbers
Buffy Coat Smear
For easier location of bacteria & parasites
Buffy Coat Smear
Buffy coat smear is considered a concentrate smear for WBCs, thus it recommended when the WBC count is _________.
Buffy Coat Smear
For blood parasites examination.
Thick Blood Smear
CHARACTERISTICS OF A PROPERLY PREPARED WEDGE SMEAR
1. Length: must occupy at least (?) of the slide
2. Should be free of (?)
3. Should have (?) appearance
4. Should be (?) than the stationary slide
5. Must have (?) from thick to thin
6. (?) to allow proper fixation
7. Should terminate in a (?)
8. Should contain at least (?) in which 50% of the RBCs do no overlap each other; the remainder ma overlap in doublets or triplets and the central pallor should be clear
1/2 to 2/3
waves, holes, and ridges
smooth and even
narrower
gradual transition
Thin enough
feathery tail
10 LPF
Too large drop of blood
TOO THICK SMEAR
Too fast spread
TOO THICK SMEAR
Too high angle
TOO THICK SMEAR
High hematocrit
TOO THICK SMEAR
Too small drop of blood
EXTREMELY THIN SMEAR
Too slow spread
EXTREMELY THIN SMEAR
Too low angle
EXTREMELY THIN SMEAR
Low Hematocrit
EXTREMELY THIN SMEAR
Accumulation of nucleated cells
GRITTY APPEARANCE
large number of leukocytes
GRITTY APPEARANCE
Use of heparin as anticoagulant
GRITTY APPEARANCE
Too slow spreading/delay in spreading
GRITTY APPEARANCE
Using only a part of the drop of blood
GRITTY APPEARANCE
Rough edge or dirty spreader slide
GRITTY APPEARANCE
METHODS OF MANUAL DIFFERENTIAL COUNT
WBCs are counted in consecutive fields as the blood film is moved from side to side
Cross-sectional/Crenellation Method
WBCs are counted in consecutive fields from the tail toward the head of the smear
Longitudinal Method
Ideal if smear is thin enough since cells can be identified all the way.
Longitudinal Method
WBCs are counted in a pattern of consecutive fields near the tail on a horizontal edge. Count 3 consecutive horiontal edge fields, 2 fields towards the center, 2 horiontal and 2 vertical to the edge
Battlement Method
Using the Oil Immersion Objective:
Estimate (?); Examine (?)
Perform WBC Differential count - A total of (?) are counted and classified according to type
Platelets and WBCs
RBC morphology
100 WBCs
The total number of cells to be counted could be decreased or increased when any of the following criteria are observed:
(?) if the total WBC count is < 1 x 10 9/L (< 1,000/µL)
Count (?) if in the first 100 count, any of the following are observed::
50 Cells
additional 100 cells
> (?) Eosinophils
(?) Basophils
(?) Monocytes
(?) are greater than Neutrophils
10%
2%
11%
Lymphocytes
METHODS OF CLASSIFYING NEUTROPHILS
WBCs are classified based on the number of lobes indication their age
ARNETHS CLASSIFICATION
- WBCs are classified according to granulation
SCHILLINGSS CLASSIFICATION
Younger forms are placed on the left; mature and old forms on the right
SCHILLINGSS CLASSIFICATION
1 round or indented nucleus
I
5% (blast cells)
I
2 nuclear divisions
II
35 %
II
3 nuclear divisions
III
41 %
III
4 nuclear divisions
IV
17 %
IV
5 or more
V
2% (oldest)
V
Schillings Hemogram
1 2 3 4 5 Eos Baso Lym Mono
Schillings Hemogram Shift to the LEFT:
increase in immature forms
Schillings Hemogram Shift to the RIGHT:
- increase in mature forms
young forms with accompanying WBC count.
Regenerative Shift to the Left
Infections, appendicitis & acute sepsis
Regenerative Shift to the Left
young forms with N or WBC count
Degenerative shift to the Left
Typhoid fever and TB
Degenerative shift to the Left
Pernicious anemia or megaloblastic anemia
Shift to the RIGHT
: cells that possess segments or lobes connected with a distinct thin chromatin filament
Filamentous
: nucleus with thinned-out portion
Non-filamentous
REPORTING
1. Relative count - Cells are reported in
2. Absolute count - Cells are reported as
% or decimal
number per volume of blood (e.g.: cells /µL or /L )
Absolute count =
Relative count (%) x total WBC count
47 77%
PM Neutrophil
1.8 7.8 x 109/L
PM Neutrophil
0 7%
Band cells
0 0.7 x 109/L
Band cells
20 40%
Lymphocytes
1.0 4.8 x 109/L
Lymphocytes
2 10%
Monocytes
0.01 0.8 x 109/L
Monocytes
0 6%
Eosinophils
0 0.6 x 109/L
Eosinophils
0 1%
Basophils
0 0.2 x 109/L
Basophils
are volatile
Circulating cells
cause lymphocytes & eosinophils to disappear from circulation within 4 8 hrs
Corticosteroid hormones
causes granulocytosis within minutes
Epinephrine
CLINICAL SIGNIFICANCE of variation in counts:
Acute Infections -
Neutrophilia Pathologic causes
bacterial, some viral, fungal, parasitic
Neutrophilia Pathologic causes
Acute Inflammatory conditions
Neutrophilia Pathologic causes
vasculitis
Neutrophilia Pathologic causes
response to tissue injury
Neutrophilia Pathologic causes
Malignancy (neoplastic growth)
Neutrophilia Pathologic causes
carcinomas
Neutrophilia Pathologic causes
sarcomas
Neutrophilia Pathologic causes
myeloproliferative disorders
Neutrophilia Pathologic causes
Tissue necrosis
Neutrophilia Pathologic causes
burns
Neutrophilia Pathologic causes
trauma
Neutrophilia Pathologic causes
MI
Neutrophilia Pathologic causes
RBC hemolysis
Neutrophilia Pathologic causes
Metabolic disorders
Neutrophilia Pathologic causes
Drugs, chemicals, venoms
Neutrophilia Pathologic causes
corticosteroids
Neutrophilia Pathologic causes
growth factors
Neutrophilia Pathologic causes
Response to therapy
Neutrophilia Physiologic causes
In response to physical or emotional stimuli
Neutrophilia Physiologic causes
stress, exercise, smoking, pregnancy
Neutrophilia Physiologic causes
Stem cell disorders
Neutropenia Inherited causes
Genetic disorder of the immune system
Neutropenia Inherited causes
Disorders of cellular development
Neutropenia Inherited causes
Intrinsic defects
Neutropenia Inherited causes
Fanconis
Neutropenia Inherited causes
Kostmanns
Neutropenia Inherited causes
Cyclic neutropenia
Neutropenia Inherited causes
Chediak-Higashi
Neutropenia Inherited causes
BM/stem cell destruction
Neutropenia Acquired causes
megaloblastic anemia
Neutropenia Acquired causes
(severe folic & Vit. B 12 deficiency)
Neutropenia Acquired causes
myelodysplasia
Neutropenia Acquired causes
marrow failure
Neutropenia Acquired causes
marrow replacement
Neutropenia Acquired causes
Infectious - any overwhelming infection
Neutropenia Acquired causes
Infectious diseases
Neutropenia Acquired causes
tularemia
Neutropenia Acquired causes
typhoid
Neutropenia Acquired causes
brucellosis
Neutropenia Acquired causes
hepatitis
Neutropenia Acquired causes
influenza, measles, mumps, rubella
Neutropenia Acquired causes
IM
Neutropenia Acquired causes
malaria
Neutropenia Acquired causes
Drugs - cancer chemotherapy, chloramphenicol, sulfas/other antibiotics, phenothiazines,benzodiazepine, antithyroids, anticonvulsants, quinine, quinidine, indomethacin, procainamide, thiazides
Neutropenia Acquired causes
Liver disease
Increase Neutrophil destruction
storage diseases
Increase Neutrophil destruction
LE Radiation
Increase Neutrophil destruction
cytotoxic drugs
Increase Neutrophil destruction
benzene
Increase Neutrophil destruction
Toxins - alcohol, benzene compounds
Increase Neutrophil destruction
Immune-mediated
Increase Neutrophil destruction
collagen vascular disorders
Increase Neutrophil destruction
RA
Increase Neutrophil destruction
AIDS
Increase Neutrophil destruction
thers - starvation, hypersplenism
Increase Neutrophil destruction
Inherited, malignant or reactive
Eosinophilia
Hypoadrenalism
Eosinophilia
Inflammatory - eosinophilic fasciitis
Eosinophilia
Allergic reaction
Eosinophilia
asthma
Eosinophilia
hay fever
Eosinophilia
drug reaction
Eosinophilia
allergic vasculitis,
Eosinophilia
serum sickness
Eosinophilia
Neoplastic diseases
Eosinophilia
Hodgkin lymphoma
Eosinophilia
CML
Eosinophilia
collagen vascular disease
Eosinophilia
ulcerative colitis
Eosinophilia
L-tryptophan eosinophilic myalgia
Eosinophilia
T cell lymphomas
Eosinophilia
Parasitism - Nematodes, Trematodes, Cestodes (trichinosis, filariasis, schistosomiasis)
Eosinophilia
Nonparasitic infections
Eosinophilia
systemic fungal
Eosinophilia
Scarlet fever
Eosinophilia
chlamydial pneumoniaof infancy
Eosinophilia
gonnorrhea
Eosinophilia
hansens disease
Eosinophilia
Associated with recovery phase of most infections during drug specific parasite manifestation
Eosinophilia
Skin disorder
Eosinophilia
psoriasis
Eosinophilia
eczema
Eosinophilia
pemphigus
Eosinophilia
dermatitis
Eosinophilia
herpetiformis
Eosinophilia
erythema multiforme
Eosinophilia
exfoliative dermatitis
Eosinophilia
Respiratory
Eosinophilia
pulmonary eosinophilic syndromes
Eosinophilia
Lofflers
Eosinophilia
tropical pulmonary eosinophilia
Eosinophilia
Churg-Strauss syndrome
Eosinophilia
Idiopathic hypereosinophilic syndromes - affecting heart, liver, spleen,CNS, other organs
Eosinophilia
certain drugs
Eosinophilia
hematologic and visceral malignancies
Eosinophilia
GI inflammatory diseases
Eosinophilia
sarcoidosis
Eosinophilia
Wiskott-Aldrich syndrome
Eosinophilia
Allergy - food, drugs, foreign proteins
Basophilia
Chronic hemolytic anemia - especially post splenectomy
Basophilia
Hypothyroidism
Basophilia
Infections - variola, varicella
Basophilia
Ulcerative colitis
Basophilia
Inflammatory diseases - collagen vascular disease, ulcerative colitis
Basophilia
Long-term foreign antigen stimulation
Basophilia
Myeloproliferative disorders
Basophilia
Systemic mast cell disease, chronic hypersensitivity states in the absence of the specific allergen
Basophilia
Estrogen therapy
Basophilia
During acute infections
Basopenia
Stress; Hyperthyroidism
Basopenia
Increased levels of glucocorticoids
Basopenia
Reactive (Immune hypersensitivity reaction)
Basopenia
Infections - many viral, pertussis, tuberculosis, toxoplasmosis, rickettsial, Whooping cough, brucellosis, sometime tuberculosis, secondary syphilis, Viral hepatitis, IM, mumps cytomegalovirus,
Lymphocytosis
Chronic inflammations - ulcerative colitis, Crohns
Lymphocytosis
Immune mediated - drug sensitivit, vasculitis, graft rejection, Graves, Sjgrens
Lymphocytosis
Hematologic - ALL, CLL, lymphoma
Lymphocytosis
Stress - acute, transient
Lymphocytosis
Destructive - radiation, chemotherapy, corticosteroids
Lymphocytopenia
Debilitative - starvation, aplastic anemia, terminal cancer, collagen vascular disease, renal failure
Lymphocytopenia
Infections - viral hepatitis, influenza, typhoid fever, TB
Lymphocytopenia
AIDS associated - HIV cytopathic effect, nutritional imbalance, drug effect
Lymphocytopenia
Congenital immunodeficiency - Wiskott-Aldrich syndrome
Lymphocytopenia
Abnormal lymphatic circulation - intestinal lymphangiectasia, obstruction, thoracic duct drainage/rupture, CHF
Lymphocytopenia
Infections - tuberculosis, subacute bacterial endocarditis, syphilis, protozoan, rickettsial
Monocytosis
Recovery from neutropenia
Monocytosis
Hematologic - leukemias, myeloproliferative disorders, lymphomas, multiple myeloma
Monocytosis
Inflammatory diseases - collagen vascular disease, chronic ulcerative colitis, sprue, myositis, polyarteritis, temporal arteritis
Monocytosis
Monocytopenia is uncommon as an isolated finding.
Monocytopenia
Following administration of glucocorticoids (During therapy with prednisone, monocytes fall during the first few hours after the first dose but return to above original levels by 12 hours)
Monocytopenia
Hairy cell leukemia (HCL)
Monocytopenia
During overwhelming infections that also cause neutropenia
Monocytopenia
