Hematology 1

Carotid arteries

Jugular veins

Arteries & Veins in the body

Head & Neck

• Carotid Arteries (Common, Internal, External) – Supply oxygenated blood to the brain, face, and neck.

• Jugular Veins (Internal & External) – Drain deoxygenated blood from the brain, face, and neck back to the heart

Aorta

subclavian arties

brachial artery

radial and ulnar arteries

pulonary arteries

Upper Body (Heart, Arms, Lungs)

Arteries:

• Aorta – The largest artery, carries oxygenated blood from the heart to the entire body.

• Subclavian Arteries – Supply blood to the arms and shoulders.

• Brachial Artery – Runs along the upper arm, commonly used for blood pressure measurement.

• Radial & Ulnar Arteries – Supply blood to the forearm and hand (radial artery is used for pulse check).

• Pulmonary Arteries – Carry deoxygenated blood from the heart to the lungs (exception: an artery carrying deoxygenated blood).

Superior Vena Cava

Subclavian veins

Pulmonary veins

Veins:

• Superior Vena Cava – Collects blood from the upper body and returns it to the heart.

• Subclavian Veins – Drain blood from the arms into the superior vena cava.

• Pulmonary Veins – Carry oxygenated blood from the lungs to the heart (exception: a vein carrying oxygenated blood).

Abdominal arota

renal arteries

lilac arteries

femoral artery

popliteal, tibial, and dorsalis pedis arteries 

Abdomen & Lower Body

Arteries:

• Abdominal Aorta – Supplies blood to organs like the stomach, liver, kidneys, and intestines.

• Renal Arteries – Supply blood to the kidneys.

• Iliac Arteries – Branch from the aorta to supply the pelvis and lower limbs.

• Femoral Artery – Main artery supplying the thigh and leg.

• Popliteal, Tibial, and Dorsalis Pedis Arteries – Supply blood to the knee, lower leg, and foot.

Inferior vena cava

hepatic vein

renal vein

iliac vein

femoral, popliteal, and tibial vein

Abdomen and lower body veins

Veins:

• Inferior Vena Cava – Collects blood from the lower body and returns it to the heart.

• Hepatic Veins – Drain blood from the liver.

• Renal Veins – Drain blood from the kidneys.

• Iliac Veins – Drain blood from the pelvis and legs.

• Femoral, Popliteal, and Tibial Veins – Drain blood from the legs and feet back toward the heart

What is the human blood composition? (amount of blood in humans, and also what precent of each component is in the blood)

Blood composition - around 7% of body weight, equal to 5L in adults

• Plasma dissolves all the cellular elements, 50-60% of blood is plasma and the rest is RBC’s 

• Plasma also contains: WBC’s, platelets, proteins, lipids, and

RBC formation, size, lifespan, and numbers

bone marrow

6-8 micrometers

100-120 days

4-8million/microlitre

WBC formation, size, lifespan, and numbers

bone marrow, lymphatic tissue

9-8 micrometers

24hrs-yrs

4.5k-11k/microlitre

Platelet (thrombocytes) formation, size lifespan, numbers

bone marrow

1-4 micrometers

8-12 days

150k-400k/microlitre

What is stem cell differentiation?

all blood cells originate from hematopoietic (rbc producing; HSCs) stem cells in the bone marrow

What are the two lines the HSCs can differentiate into?

Lineage Commitment – HSCs differentiate into either

• Myeloid lineage (producing red blood cells, platelets, and some white blood cells)

• Lymphoid lineage (producing lymphocyte; B cells, T cells, and natural killer cells

For RBCs (erythropoeisis);

what is the origin/ path?

where do they mature

final form

1. Red Blood Cells (Erythropoiesis)

Origin: Myeloid stem cells → Erythroblasts → Reticulocytes (teens)

Mature in: Bone marrow

Final form: Erythrocytes (RBCs), which carry oxygen.

For WBCs (leukopoeisis); granulocytes

what is the origin/ path?

where do they mature

final form

1. White Blood Cells (Leukopoiesis)

Granulocytes (Neutrophils, Eosinophils, Basophils) (have granules; tiny sacs with enzymes to attach intruders) - lobed nuclei to squeeze and have flexibility 

• Origin: Myeloid stem cells → Myeloblasts

• Mature in: Bone marrow

• Function: Fight infections and inflammation.

For Monocytes; (mega munchers)

what is the origin/ path?

where do they mature

final form

Monocytes (Macrophages & Dendritic Cells) (mega munchers)

• Origin: Myeloid stem cells → Monoblasts

• Mature in: Blood (monocytes) → Tissues (macrophages/eaters & dendritic cells/messengers, snitches)

• Function: Engulf bacteria and present antigens

For lymphocytes;

what is the origin/ path?

where do they mature

final form

Lymphocytes (T cells, B cells, NK cells)  (long term defence)

• Origin: Lymphoid stem cells → Lymphoblasts

Mature in:

T cells → Thymus (immunity)

B cells → Bone marrow (antibody production)

Natural Killer (NK) cells → Bone marrow (attack infected cells)

For platelets (thrombopoiesis)

what is the origin/ path?

where do they mature

final form

1.  Platelets (Thrombopoiesis)

Origin: Myeloid stem cells → Megakaryoblasts

Mature in: Bone marrow

Final form: Platelets (Thrombocytes), which help in blood clotting

What are some growth factors?

Erythropoietin

Thrombopoietin

Colony- Stimulating Factors (think army)

Growth factors:

• Erythropoietin (EPO) – Helps make red blood cells (RBCs), which carry oxygen

• Thrombopoietin (TPO) – Helps make platelets, which stop bleeding by forming clots

• Colony-Stimulating Factors (CSFs) – Help make white blood cells (WBCs), which fight infections

Define each RBC disorder

anemia

hyperslenism

sickle cell disease

polycthemic vera (hint; POLY)

Red Blood Cell Disorders

• Anemia – Low red blood cell count, leading to fatigue and weakness

• Hypersplenism - spleen is overactive in blood cell removal 

• Sickle Cell Disease – Abnormally shaped red blood cells that block blood flow = pain

• Polycythemia Vera – Too many red blood cells, making the blood thick, increase clot risk

What does hematological disease effect?

blood, bone marrow, and lymphatic system

Define each WBC disorder

leukemia

lymphoma - give a name too (hint; Hod)

neutropenia

White Blood Cell Disorders

• Leukemia – Cancer of white blood cells (grow uncontrollably), affecting immune function 

• Lymphoma – Cancer of the lymphatic system (eg; Hodgkin’s and non-Hodgkin’s lymphoma)

• Neutropenia – Low neutrophil count, increasing infection risk

Define each platelet disorder 

thrombocytopenia

thrombocythemia

Platelet Disorders

• Thrombocytopenia – Low platelet count, causing excessive bleeding.

• Thrombocythemia – Too many platelets, leading to clotting issues.

Define each plasma disorder

hemophilia A & B

Von Willebrand Disease

Multiple Myeloma

Plasma Disorders

• Hemophilia – A bleeding disorder due to missing clotting factors; Factor VIII (Hemophilia A) or Factor IX (Hemophilia B)

• Von Willebrand Disease – A genetic disorder caused by a missing/defective protein (von willebrand factor), affecting blood clotting

• Multiple Myeloma – Cancer of plasma cells in the bone marrow (abnormal plasma cells grow uncontrollably, leads to faulty antibodies), weakens bones and reduces immunity 

what are inherited and acquired disorders?

Inherited 

Eg; hemophilia; bleeding problems from defective coagulation factors 

Acquired

Eg: viral infections, medication, radiation, heart conditions

How is specimen picked for hematology lab testing?

• Specimen - capillary and venous blood samples, choice made is based on volume needed and specific biomarkers measured

What is a CBC?

• Complete blood count (CBC) - common test, analyzes numerous markers; cell counts, hemoglobin, hematocrit, platelets, blood cell morphology 

What is hemoglobin?

What % of RBC protein content does hemoglobin make up, and what is the structure of it 

Hemoglobin is an iron-rich protein in red blood cells that transports oxygen from the lungs to the body's tissues and carries carbon dioxide from the tissues back to the lungs.

Hemoglobin characteristics - make up 98% of RBC protein content and provides red colour

Structure: heme and globin groups

• Globin = quaternary structure of 4 protein chains linked together

• Heme = iron that allows the binding of oxygen

Hemoglobin reference ranges for

newborn

children

adult males

adult females

age/gender	g/dl (grams per deciliter/tenth of a L)	Conversion factor	g/l (SI units)
Newborn	16-23	x10	160-230
Children	10-14	x10	100-140
Adult males	13-17	x10	130-170
Adult females	12-16	x10	120-160

How does the specific gravity technique for hemoglobin measurement work?

Specific gravity technique (approximation)

• Involves copper sulfate solution w/specific gravity of 1.052 (approx the same as blood with a low hemoglobin concentration)

• If a drop of blood falls through = healthy hemoglobin count

• If a drop floats = low hemoglobin count

How does the clinical method, or cyanmethemoglobin measurement work for hemoglobin?

Chemical methods (cyanmethemoglobin measurement)

• Blood is reacted w/Drabkin’s solution, then reacts w/hemoglobin to form cyanmethemoglobin

• Changes the colour of the solution, then is photometrically analyzed to determine the hemoglobin concentration

Photometrically = light measurement, how much light the sample absorbs at a specific wavelength 

How does the hematocrit test measurement work for hemoglobin?

Hematocrit - test used to determine volume of patient’s RBC volume (oxygen carrying capacity)

1. Microhematocrit - test involving centrifuging a small sample of blood

• Components of blood will separate by specific gravity 

• After centrifuging - bottom of tube = RBCs, top will have WBCs, buffy coat, platelets, and plasma (order is called packed cell column)

• Specific gravity = ratio of density of a substance to the density of a reference substance (usually water at at 4°C, where water has its maximum density of 1 g/cm³)

  Formula = Density of Substance​/Density of Water (1 g/cm3)

What are the hematocrit reference values affected by?

Hematocrit reference values - affected by physiological and pathological factors

• Normal values vary by age and sex

• anemia/bleeding = lower levels

• dehydration = higher values (bc when water loss, volume of plasma decreases (liquid part of blood), but the number of RBCs stay the same = higher concentration)

What are the hematocrit reference values for

adult male

adult female

children

newborn

1yr

6yrs

What can the appearance of packed cell colulmn indicate?

Appearance of packed cell column can indicate conditions

Eg; opaque plasma = high fat levels

What are the 3 steps to performing microhematocrit tests

Performing microhematocrit tests

1. Collecting specimen

• Sample is obtained from a capillary puncture or from venus blood (w/anticoagulant)

• Blood samples should be drawn into capillary tubes through capillary action

2. Centrifuging 

• Capillary tubes must be balanced

• Generally spun at 10,000 RPM for 2-4 minutes

3. Reading test values

• Microhematocrit readers are used to determine test values

• RBC portion of tube is measured and a value is produced 

What is hematocrit and microhematocrit?

Hematocrit is a measure of the proportion of your blood that is made up of red blood cells. It's expressed as a percentage and indicates how much of your blood's volume is red blood cells compared to the liquid plasma, white blood cells, and platelets. A low hematocrit can signal anemia (not enough red blood cells), while a high hematocrit can indicate conditions like dehydration or too many red blood cells (polycythemia)

Microhematocrit is a laboratory procedure that measures the volume percentage of red blood cells in a blood sample.

What is a hemocytometer?

Hemocytometer & blood cell counting 

Blood count - automated counters and hemocytometer (microscopically)

Hemocytometer - precision made microscope slide that includes a counting chamber

• Allows manual counting of blood cells in a sample

• Typically glass slide w/2 counting chambers

• Coverglass is needed on top of the slide to confine blood sample for microscopy 

What does a moat do?

prevents overflowing

How is the hematocrit test divided?

Counting chambers - 2 on a hemocytometer, each divided into 9 squares

• Each square is divided into further segments; corners are split into 16 squares and the center is split into 625 squares

• 4 corners are for counting WBCs (larger), center used for RBCs (smaller)

What is a hemocytometer and how do u count it?

Hemocytometer - manually count cells under a microscope

Preparing sample

• Must be diluted depending on cell type

• RBCs - 1:200 dilution

• WBCs & platelets - 1:100 or 1:20 dilution

• Employees must be trained on donning PPE and items

Loading hemocytometer - micropipette around 10uL of blood into counting chamber in a smooth unbroken stream 

Cell counting pattern 

• count upper or left in a left→ right squiggle down alternating

What is the formula for calculating cell counts?

Calculating cell counts 

Formula:

cells/uL = # counted cells (DF) / a(mm2)x0.1

DF = dilution reciprocal

What is the purpose of blood smears?

Blood smears - staining and observing RBCs, WBCs and platelets microscopically

• Altered cell morphology can inform physicians about diseases; IM, sickle cell anemia, malaria

What specimen should you collect for blood smears?

Preparing blood smears

Specimen collection 

• Capillary blood is recommended 

• Venous blood can be used w/coagulant 

How do u make, preserve, and qualities of a good smear

Making the smear 

Two-slide method: 

1. Drop of blood placed on 1 end of the slide

2. Another slide edge is placed at an angle to specimen slide

3. Edge is moved into a blood sample & when the specimen spreads along the edge, quickly push the slide forward to distribute the blood 

Preserving the smear - immerse in methanol for 30-60 seconds

• Methanol is a fixative that prevents deterioration of cellular components

Qualities of a good smear

• Should cover ¾ of the slide and have gradual transition from heavy to light 

• No holes/ridges

• Cells should be evenly distributed under microscope w/o overlapping RBCs in the thin section

What does staining the smear do? Explain wrights stain procedure

Wright’s stain procedure:

1. Quickstain - dip blood smear slide into Wright’s stain, creates adequate results in 2-5 minutes

2. Two step method - smear slides placed on a staining rack and are flooded with Wright’s stain

• Then flooded w/buffer that accentuates the stain colours

3. Automatic stainer - staining machines, provide better results

What are common buffers in blood staining (resists ph change)

Common Buffers in Blood Staining: (proper pH 6.8-7.2)

• Phosphate buffer (pH 6.8–7.2) – Used in Wright's, Giemsa, and Romanowsky stains

• Sørensen’s buffer – A phosphate buffer used in hematology

What are different types of blood parts used for and why?

whole blood

plasma

serum

capillary

venous

arterial

Blood Type	Used For	Why
Whole Blood	CBC, blood cultures, immunology tests	Contains all components (RBCs, WBCs, platelets, plasma).
Plasma	Coagulation tests (e.g., PT, aPTT), electrolytes, hormones	Liquid part of blood contains proteins, hormones, nutrients, and waste.
Serum	Blood chemistry, liver enzymes, antibody testing	Blood after clotting, ideal for assessing electrolytes and antibodies
Capillary Blood	Blood glucose monitoring, newborn screening	Convenient for quick results, especially for glucose tests.
Venous Blood - most popualr	Routine tests (CBC, liver/kidney function, cholesterol)	Standard source for most blood tests, easy to collect in larger volumes.
Arterial Blood	Arterial blood gas (ABG) analysis	Provides accurate oxygen, carbon dioxide levels for respiratory and metabolic function.

What is normal RBC morphology?

shape, size, color, nucleus, function

 Red Blood Cells (RBCs):

• Shape: Biconcave discs (donut-shaped, but without a hole in the center).

• Size: About 7-8 micrometers in diameter.

• Color: Pinkish due to hemoglobin content.

• Nucleus: None, do not have a nucleus in their mature form.

• Function: Carry oxygen and carbon dioxide.

What is normal platelet morphology?

shape, size, color, nucleus, function

 Platelets:

• Shape: Disc-shaped with a slightly irregular outline.

• Size: About 2-3 micrometers in diameter.

• Nucleus: None, as they are fragments of larger cells called megakaryocytes (large bone marrow cells)

• Function: Play a key role in blood clotting by forming plugs in response to injury

What is normal neutrophil morphology?

shape, size, color, nucleus, function

White Blood Cells (WBCs):

Lobed: allows easier movement and performing specialized immune functions, eg squeezing through tiny blood vessels or engulfing pathogens

Granules: attack stuff and do the function

Neutrophils:

• Shape: Multi-lobed nucleus (3-5 lobes).

• Size: About 12-15 micrometers

• Color: Pale cytoplasm with fine granules.

Function: First responders to infections, especially bacterial.

what is normal lymphocytes morphology?

shape, size, color, nucleus, function

• Shape: Large round nucleus with a thin rim of cytoplasm.

• Size: About 7-10 micrometers.

• Color: Dark blue nucleus with a light blue cytoplasm.

Function: Important for adaptive immunity, producing antibodies (B cells) and attacking infected cells (T cells)

What is normal monocyte morphology?

shape, size, color, nucleus, function

Monocytes: EATERS AND SNITCHES

• Shape: Kidney-shaped or horseshoe-shaped nucleus.

• Size: About 15-20 micrometers.

• Color: Pale blue cytoplasm.

• Function: Differentiate into macrophages or dendritic cells (snitches) to fight infection and clear debris.

What is normal eosinophil morphology?

shape, size, color, nucleus, function

Eosinophils:

• Shape: Bi-lobed nucleus (often resembling a figure 8).

• Size: About 12-17 micrometers.

• -orange granules in cytoplasm.

• Function: Involved in allergic reactions and combating parasitic infections

What is normal basophil morphology?

shape, size, color, nucleus, function

Basophils:

• Shape: Bi-lobed nucleus, often obscured by granules.

• Size: About 10-15 micrometers.

• Color: Dark purple granules in the cytoplasm.

• Function: Release histamine during allergic reactions and inflammation

What does histamine do?

Histamine - causes blood vessels to expand and become more permeable (allowed substances to pass through)

Image of all wbcs

What is Anisocytosis?

Anisocytosis - mix of RBC sizes in blood 

What are microcytic and macrocytic rbcs caused by?

Anisocytosis - mix of RBC sizes in blood 

1. Microcytic RBCs = smaller

• caused by iron deficiency anemia, thalassemia (genetic blood disorder; lack of hemoglobin)

2. Macrocytic RBCs = larger

• Caused by vitamin B12 deficiency, alcoholism, liver disease, folic acid deficiency (type of vitamin B9)

What is Poikilocytosis and what conditions is it seen in?

Poikilocytosis (genetic abnormalities in morphology)

• Seen in anemia, liver disease genetic blood disorders

What are drepanocytes? (cause and effect)

1. Sickle Cells (drepanocytes):

• Shape: Crescent or sickle-shaped (in low oxygen conditions)

• Cause: Due to sickle cell anemia, where the hemoglobin in RBCs is abnormal 

• Effect: RBCs become rigid and sticky, blocking blood flow, insufficient oxygen supply for organs, and causing pain

What are spherocytes? cause and effect

1. Spherocytes:

• Shape: Spherical (round) 

• Cause: Seen in conditions like hereditary spherocytosis or autoimmune hemolytic anemia (destroyed by spleen when they get trapped)

• Effect: These cells are more prone to destruction in the spleen, leading to anemia

What are elliptocytes? cause and effect

1. Elliptocytes (ovalocytes):

• Shape: Oval or elliptical

• Cause: Often seen in hereditary elliptocytosis (destroyed by spleen)

• Effect: These cells are less flexible, which may lead to mild hemolysis (destruction of RBCs) and anemia.

What are stomatocytes? cause+effect

1. Stomatocytes:

• Shape: Oval with a central slit-like area (mouth-shaped)

• Cause: Associated with hereditary stomatocytosis or liver disease

• Effect: Can lead to membrane instability and impaired oxygen transport.

What are target cells? cause+ effect

1. Target Cells:

• Shape: RBCs with a central area of hemoglobin surrounded by a clear ring (like a target)

• Cause: Found in conditions like thalassemia, liver disease, and iron deficiency anemia

• Effect: RBCs may have reduced oxygen-carrying ability

What are keratocytes? cause+effect

1. Keratocytes:

• Shape: RBCs with blister-like vesicles or spiky projections

• Cause: Often a sign of mechanical damage to RBCs (burns, movement through artificial heart valve, cut by fibrin strand) or conditions like microangiopathic hemolytic anemia

• Effect: These cells are fragile and may be destroyed prematurely

What are crenated cells?

1. Crenated Cells: improper blood smear procedure

• Shape: RBCs with spiny projections or irregular edges

• Cause: Can result from dehydration, where the cell loses water and shrinks or prolonged exposure to anticoagulant 

• Effect: These cells are less flexible and can have impaired circulation

What are nonchromic, hypochromic, and hyperchromic cells

Normochromic cells - RBCs w/correct hemoglobin amount 

Hypochromic = lower hemoglobin content, very thin outer rim

Hyperchromic = higher hemoglobin content, no central divot that normal RBC’s have 

What are rbc inclusions? and what are they caused by

RBC Inclusions - abnormal structures/structures inside RBC’s

• Diseases caused by toxicity, hemolytic anemia, vitamin deficiency

What is the appearance, cause, conditions, and effect of basophilic stippling?

1. Basophilic Stippling:

• Appearance: Small, blue granules seen in the cytoplasm of red blood cells

• Cause: granules are remnants of ribosomal RNA (or ribosomes) that are not removed during cell maturation

• Associated Conditions: Lead poisoning, thalassemia, sideroblastic anemia (can't properly use iron to make hemoglobin), chronic liver disease

Effect: often a sign of impaired RBC maturation or a disturbance in hemoglobin production

What is the appearance, cause, conditions, and effect of Howell-jolly bodies?

 Howell-Jolly Bodies:

• Appearance: Small, round, purple or dark blue inclusions in the RBCs, representing nuclear remnants.

• Cause: These bodies are left over from the nucleus when RBCs lose it during maturation

• Associated Conditions: Splenectomy (removal of the spleen), hemolytic anemia, megaloblastic anemia (due to vitamin B12 or folate deficiency)

• Effect: The spleen normally removes these remnants, so their presence suggests the spleen is not functioning or absent

What is the appearance, cause, conditions, and effect of cabot rings?

. Cabot Rings:

• Appearance: Ring-shaped or figure-eight structures in the cytoplasm of RBCs, often purple.

• Cause: They are believed to be remnants of the mitotic spindle or nuclear material

• Associated Conditions: Severe anemia (eg megaloblastic anemia; abnormally large RBCs that don't work properly), lead poisoning, myelodysplastic syndromes (group of bone marrow disorders)

• Effect: These are rare but indicate disrupted cell maturation or abnormalities in RBC production

What is the appearance, cause, conditions, and effect of nucleated rbcs?

. Nucleated RBCs:

• Appearance: RBCs that still contain a nucleus

• Cause: immature RBCs are being released into the bloodstream prematurely, happens when the bone marrow is under stress or producing RBCs rapidly

• Associated Conditions: Severe anemia, bone marrow disorders, hemorrhage (bleeding), hypoxia (insufficient oxygen supply to body tissues)

• Effect: Normally, RBCs lose their nucleus in the bone marrow before entering circulation, presence in the blood suggests abnormal bone marrow function or increased RBC production

What is leukopenia and the cause?

Leukopenia - low WBC count 

Causes: infections, antibiotics, chemotherapy, radiation

Bone marrow disorders - leukemia, aplastic anemia, myelodysplastic syndrome

What is leukocytosis and the cause?

Leukocytosis - high WBC count

Causes: infections, exercise, anxiety, medications, steroids

what is leukemias?

Leukemias - unrestrained production of leukocytes in bone marrow

• Inhibits growth of other cells, leading to anemia

What is cause of thrombocytopenia and thrombocytosis?

Thrombocytopenia - low levels caused by radiation, certain drugs, alcoholism, leukemia

Thrombocytosis - high levels caused by inflammatory conditions, reaction to other blood disorders