Chapter 1 – An overview of Clinical - sodium dodecyl sulfate (SDS): used

Laboratory Hematology instead of cyanide to reduce toxicity

• Average human possesses 5L of blood Hematocrit (Hct)

• Blood fxns: - ratio of RBC volume and whole blood
- transports O2 from lungs to tissues volume
- clears CO2 in tissues - aka packed cell volume
- transports glucose, proteins, fats - packed cells = RBCs
- moves wastes to the liver and kidneys
• Plasma – transports and nourishes blood Buffy coat
• 3 families of blood cells: - layer between RBCs and plasma
- RBCs (erythrocytes) - contains WBCs and platelets
- WBCs (leukocytes)
- Platelets (thrombocytes) To compute for RBC indices, 3 numerical
results may be used:
HISTORY 1.) Hb
Athanasius Kircher (1657) 2.) Hct
- “worms” in the blood 3.) RBC count
Anton van Leeuwenhoek (1674)
- account of RBCs Mean Cell Volume (MCV)
Giulio Bizzozero (late 1800s) - RBC diameter on Wright stained film
- platelets as “petite plaques” - Variation: anisocytosis
James Homer Wright (1902)
- Wright stain Mean Cell Hemoglobin Conc. (MCHC)
- Wright’s Romanowsky type stain - RBC staining intensity/ degree of pallor
(polychromatic, a mixture of acidic and - Variations: anisochroma/ anisochromasia
basic dyes)
Mean Cell Hemoglobin
RBC morphology – cell appearance (cell color, - mass of hemoglobin
size, shape, cytoplasmic inclusions and nuclear - reflects MCHC
condensation)
Red Cell Distribution Width (RDW)
RED BLOOD CELLS - degree of variation in RBC volume
- Anucleate biconcave cells with Hb - related to size
- Pink to red - variations: poikilocytosis
- 6 – 8 um diameter
- one-third zone of pallor at the center RETICULOCYTES
- Anemia (↓ RBC count): loss of O2 carrying - young RBCs/ polychromatophilic
capacity erythrocytes
- Polycythemia (↑ RBC count): leads to - production site: bone marrow
hyperviscosity - exceed 6-8 um
- 1:200 dilution - stain slightly blue-gray
- diluting fluid: 0.85% NSS - blood loss and certain anemias: indicate
bone marrow regeneration
Coulter counters - Methylene blue dyes (nucleic acid stains/
– first electronic counter to count RBCs vital stains)
– Coulter principle of direct electrical - contain RNA
impedance - absolute reticulocyte count
– Joseph and Wallace Coulter - immature reticulocyte count/ immature
reticulocyte fraction: sensitive measure of
Hb, Hct and RBC indices bone marrow fxn
Hemoglobin (Hb)
- Drabkin rgt ( pot. Cyanide, pot. WBCs
Ferricyanide) - not really blood cells
- 540 nm wavelength - protecting their host from injury and
infection
- “hitch a ride” (usually from bone marrow - recognize foreign antigens
or lymphoid tissue) to their destination - mount antibody (humoral) and cell-
- nearly colorless in an unstained cell mediated antagonistic responses
suspension - round featureless nuclei
- chronic leukemia (↑ WBC count): milky - thin rim of ningranular cytoplasm
appearance - lymphocytosis (↑): viral infections
- counted visually by microscope, - lymphopenia/lumphocytopenia (↓):
hemacytometer, Thoma pipet imunodefiecy/ long term drug
- 1:20 dilution administration
- diluent: dilute acetic acid in NSS - chronic lymphocytic leukemia: older than
o acid causes RBCs to lyse; w/o it RBCs 70
would obscure WBCs - acute lymphoblastic leukemia: childhood
- leukopenia (↓ WBC count); leukocytosis (↑ leukemia
WBC count)
Monocytes (monos)
TYPES OF WBCs - immature macrophages
Polymorphonuclear nuetrophils (PMNs, - most abundant cell in the body
segmeneted neutrophils or segs) - occupy every body cavity
- engulf and destroy bacteria - motile and immobilized
- neutrophilia (↑): bacterial infection - phagocytosis
- neutropenia (↓): long-term drug - assist lymphs in presentation of
administration/ viral immunogenic epitopes
- gray cytoplasm; lobulated nucleus
Band neutrophils (bands) - monocytosis (↑): leukemia
- less mature than segs - monocyte-macrophage cell line: source for
- left shift (↑): bacterial infection chronic and acute monocytic leukemia

• cytoplasm of segs and bands: contains PLATELETS (plts)

pink-staining granules filled with - true blood cells
bactericidal secretions - maintain vessel integrity by vessel wall
repairs
Eosinophils (EOs) - process:
- bright orange 1.) adhere to damaged bld. Vessels
- regular cytoplasmic granules filled with 2.) aggregates w neighboring platelets to
antihistamine plug the vessels
- eosinophilia (↑): allergy or parasitic 3.) secrete enzymes that trigger
infection thrombosis or clot formation
- control hemostasis (seals wounds)
Basophils (basos) - maintains vascular patency
- dark purple - 2-4 um in diameter
- irregular cytoplasmic granules that obscure - round/oval; anucleate; slightly granular
nucleus - counting is confined in the center square
- basophilia (↑): hematologic disease (e.g. millimeter of hemacytometer
leukemia) - phase microscopy
- thrombocytosis (↑): inflammation or
• Segs, bands, EOs, Basos: granulocytes trauma
(prominent cytoplasmic granules) - essential thrombocythemia (extremely ↑
• Leukemia: uncontrolled proliferation of and uncontrolled plt production):rare
WBC malignant condition
o Chronic myelogenous (granulocytic) - thrombocytopenia (↓):drug treatment w
o Acute myeloblastic easy bruising and uncontrolled hemorrhage
• Eosinopenia and basopenia: theoretical
and unused COMPLETE BLOOD COUNT (CBC)
- short draw: inappropriate anticoaguant-to-
Lymphocytes (lymphs) specimen ratio
- host immunity
- specimen accession: registers the - uses graduated conc. of saline soln’s to
specimen in the work list detect spherocytes (proportionally reduced
- flag: abnormal results in automated surface membrane area)
profiling instruments - in hereditary spherocytic or warm
- reflex blood film examination: performed autoimmune hemolytic anemia
when there is a flag
Glucose 6-phosphate dehydrogenase asay
BLOOD FILM EXAMINATION - tests for inherited RBC enzyme deficiency
- “wedge-prep” method (dries, fixes and causing severe episodic hemolytic anemia
stains using Wright or Wright-Giemsa)
- WBC differential: reviews, identifies and Sickle cell solubility screening assay
tabulates 100 WBCs to determine their % - follow up test: Hb electrophoresis
distribution - detect sickle cell anemia, thalassemia,
qualitative Hb abnormalities
ENDOTHELIAL CELLS
- structural
- do not flow in the bloodstream
- endodermal cells that form the inner
surface of blood vessels
- maintains normal blood flow
- snaring plts in times of injury’
- enabling WBCs to escape
- secretions: cytokines

COAGULATION
- coagulation system:
o complex sequence of plasma proteins
to produce clot formation after vessel
injury
o 6-8 enzymes exert control over
coagulation mechanism
o third system of enzymes an cofactors
digest clots and restore vessel patency
(fibrinolysis)

ADVANCED HAMATOLOGY PROCEDURES

Bone marrow aspirates and biopsy specimens
- used to analyze nucleated precursor of
RBCs
- erythroid: RBC series
- myeloid: bands, segs, EOs, basos
- meakaryocyte: plt
- myeloperoxidase, Sudan black B, PAS,
nonspecific and specific esterase, tertrate
resistant acid phosphatase and alkaline
phosphatase: cytochemical stains used to
differentiate abnormal myeloid, eryhtroid
and lymphoid cells

Flow cytometers (FCT)

- clinical FCT: quantitative
- laser-based FCT: qualitative

ADDT’L HEMATOLOGY PROCEDURES

Osmotic Fragility Test (OFT)