Lecture 1: METHODS, ERRORS, HEMATOPOIESIS, RBC REVIEW

- CBC (complete blood cell counting)
- WBC differential using a peripheral blood smear/film

- Flowcytometry (for immunophenotyping)
- Molecular diagnostics (gene mutations)
- Cytogenetics (chromosomal changes/translocations)
- (sometimes) cytochemical stains (MPO, SBB, NSE, etc.)
ADDITIONAL
- G6PD phenotyping
- SDS-PAGE (E.G., for hereditary spherocytosis or vWD)
- HPLC (e.g, for SCD)
- Body fluid analysis (cell count)

types of specimens
AC of choice
Factors related to phlebotomy
Handling (Storage)
Short draw effects
Over exposure to EDTA

Romanowsky stains which are polychromatic (contain eosin Y and methylene blue)

Basic and stains acidic cellular components (RNA, DNA, basophil granules) from blue-grey to dark blue in color

acidic and stains basic cellular components (hemoglobin and eosinophilic granules) from light to deep pink to red or orange in color

neutrophilis have cytoplasmic granules with neutral pH and pick up some staining characteristics from both stains (thiazine-eosinate complex).
-Neutrophil granules appear pink/violet (lilac) compared with eosinophils

Check staining quality, smear prep quality
Check sample integritiy (PLT clumps, fibrin strands, cell distribution,...)
Do WBC and PLT estimates (to find correlation with analyzer report in case of flags)
Do WBC differential
Look for abnormal WBCs (blasts) or reactive lymphs
Check alterations in RBC and PLT morphology (shape, color, size, distribution pattern)
Look for parasites, bacteria

A: Pronormoblast
B:
C:
D:
E: Retics
F: Polychromatic erythrocytes
G: Erythrocytes -RBCs
H: RBCs

A: Pronormoblast
B:
C:
D:
E:
F: Nursing cells; Erythrocyte blast island surrounding a macrophage

1. Glycolysis (Embden-Meyerhof pathway)
-Main way RBCs make energy (ATP).
-Breaks down glucose into smaller molecules to release energy.
2. Shunts (side pathways)
Hexose monophosphate pathway (HMP): Makes NADPH, which protects RBCs from damage caused by oxidation (like a built-in antioxidant system).
Rapoport-Luebering pathway: Helps RBCs control oxygen delivery to tissues by producing 2,3-BPG, which lets hemoglobin release oxygen more easily.
Methemoglobin reductase pathway: Converts methemoglobin (which can’t carry oxygen) back into hemoglobin so it can carry oxygen again.

A1: It allows RBCs to squeeze through small capillaries without breaking.

A2: Transmembrane (integral) proteins and cytoskeletal (peripheral) proteins.

A3: Band 3, GLUT-1, Rh proteins, ion pumps.

A4: Ankyrin, spectrin, actin, protein 4.1.

A5: It aids in gas exchange and anchors the cytoskeleton to the membrane.

A6: Hereditary Spherocytosis (HS).

A7: Hereditary Elliptocytosis (HE).

A8: Provides elasticity and flexibility to the RBC membrane.

A1: 95%.

A3: Globin chains and heme groups.

A4: Globin genes.

A5: Fe²⁺ (ferrous iron) + protoporphyrin IX.

A6: Four.

A7: It binds oxygen in the lungs and releases it in tissues.

A8: Four.

A: NORMAL SITUATION (STEADY STATE)
B: ALKALOSIS, LOW 2,3-DPG, HbF
C: acidosis, high temperature, high 2,3-DPG

hemoglobin holds oxygen tighter.

hemoglobin lets go of oxygen

Normocytic normochromic

stomatocytes

Ecchinocytes

Acanthocytes

Spherocytes

Schistocytes

Sickle cells

Ovalocytes / pencil cells

Codocytes aka target cells

Dacryocytes aka tear drop cells

Bite cells aka "degmacyte" (abdi said)

Microcytes

Normocytes

Macrocytes

Anisocytosis or dimorphism

Hypochromia and microcytes

Target cells

Reticulocytes; increase polychromatic

Acanthocytes and nRBCs

Schistocytes

Echinocytes

Bite cells

Basket cells aka blister cells aka purse cells

Spherocytes

Target cells in hemoglobinopathies

hgb C crystals

Schistocytes

Ovalocytes

Tear drop cells or dacrocytes

Stomatocytes

Rouleux

agglutination

holly jowell bodies

Basophilic stippling

Pappenheimer bodies

Pappenheimer bodies with prussian blue

Golf balls

Dacrocytes , macrocytes, microcytes