Hematology Unit 1

Hemoglobin Assay

-(Hgb) -Read at 540 nm on a spectrophotometer -Automation uses a modified method

Medullary/Myeloid Phase:

-Bone Marrow as the primary site -Medulla or inner part -6 months gestation -Myeloid to Erythroid ratio 3:1 FOR AN ADULT IS 2:1

Medullary phase detectable levels:

-Erythropoietin (EPO) -Granulocyte colony stimulating factor -Hgb, F, A2, and an adult A can be detected

Reticulocytes

-Immature RBC's that contain RNA -Normally make up 1-2% of RBC's -Slightly larger in diameter -Stained blue grey -Indicate bone marrow regeneration (ANEMIA)

Causes of Macrocytosis

-Increased number of reticulocytes -Antiviral treatment of HIV infection

Hepatic phase

-Liver is the primary site -Begins at about 4-5 weeks gestation -Extravascular (Liver and Spleen) -Lymphocytes for the kidney, thymus, and spleen -Hgb F, A, A2

Blood Film Examination

-Preparation: Make a wedge prep blood film on a glass microscope slide -Estimates: Visual estimates of platelets and WBC counts using a microscope -Cell morphologic form and structure -Count 100 WBC's and classify each type

Hematocrit

-Ratio of volume of RBC's to the volume of whole blood -Normal ratio is 40-50% -Packed Cell Volume (PCV) -Can be analyzed by centrifugation

Reflux Testing

-Stems from initial abnormal testing -Follow up testing due to the results of screening tests -Lab must design reflex testing protocols for common diseases

Mesoblastic phase

-Yolk Sac/AGM -18 days to 2 months gestation -Intravascular -Embryonic Hgb

A low platelet count can represent:

1. Low thrombopoiesis 2. Platelet destruction or consumption

MCV is used to classify cells as:

1. Normocytic 2. Microcytic (Cells appear smaller) 3. Macrocytic (Cells appear larger)

Components of Laboratory Testing

1. Pre-examination 2. Examination 3. Post-examination

At what point during fetal development does hematopoiesis?

18 days

Intraembryonic Hematopoiesis

4 weeks of gestation Begins in the aorta-gonads-mesonephros region Lumen of the developing aorta Definitive erythropoiesis begins!

How do we do WBC and PLT Estimates?

40 x Objective Lens

If 95% of normal individual test values value within the range then...

5% can be normal and not in the normal reference range

What percentage of the body is plasma?

55%

How much blood does a normal adult have?

6 Liters (about 7-8% of your body weight)

When does production switch over to only bone marrow?

9 months- birth

Agglutination vs. Rouleaux

Agglutination: Cluster like grapes, cold, important to keep it warm Rouleaux: Coin, Saline Solution

Rouleaux

Alignment of RBC's on top of one another Resembles a stack of coins Seen with an increase in fibrinogen to globulins Can also be an artifact (EDTA) Disappears when cells are suspended in saline

Bone marrow assays

Analyze immature blood cell precursors

Cabot Rings

Appear as a figure 8, ring or incomplete ring, stain reddish violet

At 6 months gestation, what are the sites of the hematopoiesis?

Bone Marrow and Liver

What tissues are considered primary lymphoid tissues?

Bone marrow, thymus

Causes of Microcytosis

Depending upon the population studied -Reduced iron availability -Reduced heme synthesis -Reduced globin production

Cytochemical Stains

Differentiate abnormal myeloid, erythroid, and lymphoid cells

Purple Tube Top

EDTA

Red Blood Cells

Erythrocytes Carry Oxygen Contain hemoglobin which transports O2 and CO2

What cells does the yolk sac make?

Erythrocytes (RBC's) to transport oxygen and Macrophages

Where does embryonic hematopoiesis occur?

Extraembryonic location in the YOLK SAC

Schistocyte (Cut)

Fragments of red cells, variety of shapes

Chain of Embryonic Hemoglobins

Gower 1, Gower 2, and Portland

Myeloblasts are also known as:

Granulocytes

Hct Formula

Hct= RBC(MCV)/1

Hematopoiesis

Hemato= blood -Poiesis= to make

What does Hematology mean?

Heme= Blood -ology= Study of Study of Blood

Otitis

Inflammation of the ear

Plasma Composition

Ions, Proteins, Acts as a transport medium for cell nutrients

Lymphocytes are in higher abundance in:

Kids

White Blood Cells

Leukocytes Fight against pathogens Defense against foreign antigens like bacteria and viruses

What is plasma made of?

Liquid component: water, ions, proteins, carbs, fats, enzymes Cellular elements: Erythrocytes (RBC's), leukocytes (WBC's), platelets (Plt)

Primitive Embryonic Erythroblasts

Located in the yolk sac (blood islands!) Produce embryonic hemoglobin Relate to the development of the endothelium (blood vessel lining)

MCH Formula

MCH= Hgb/RBC (10)

MCHC Formula

MCHC= Hgb/HCT (100)

MCV is greater that 100; MCHC is 32-36

Macrocytic; Normochromic

Monocytes with tissue turns into:

Macrophage

QA (Quality Assurance)

Making sure everything is the same each time

MCH

Mean Cell Hemoglobin Reflects hemoglobin mass

MCHC

Mean Cell Hemoglobin Concentration Reflects RBC staining intensity or pallor

MCV

Mean Corpuscular Volume Reflects the RBC diameter on a Wright stained smear

RBC indices

Mean Corpuscular Volume (MCV), Mean Corpuscular Hemoglobin (MCH), Mean Corpuscular Hemoglobin Concentration (MCHC)

Three Phases of Hematopoiesis:

Mesoblastic (Yok sac, AGM), Hepatic (Liver and Spleen), Medullary (Bone Marrow)

MCV less than 80; MCHC is less than 32

Microcytic; Hypochromic

Monoblasts are known as:

Monocytes

Stomatotype (Mouth)

Mouth cell, cup form, mushroom cap

Mature Red Blood Cells

Nonnucleated biconcave discs 6 to 8 microns in diameter Move wastes from the tissues to the liver and kidneys

MCV is 80-100; MCHC is 32-36

Normocytic; Normochromic

Cold Agglutinins

Not really common unless cold IgM antibodies directed against RBC antigens RBC's agglutinate and form irregular clusters

Pathologic Changes

Occur as the result of a disease or injury

Molecular Testing

Philadelphia Chromosome

What is the difference between serum and plasma?

Plasma has clotting factors, serum does not

Megakaryoblasts are also known as:

Platelets

Primitive vs. Definitive Hematopoiesis

Primitive- Blood island/yolk sac Definitive- inside the embryo/aorta/AGM

Complete Blood Count (CBC)

Quantifies the WBC's, RBC's, Hemoglobin (HgB), Hematocrit (HCT), and Platelets

Flow Cytometry

Quantitative: gives a number or % Qualitative: Presence or Absence

The rule of 3

RBC (3) = Hgb Hgb (3)- HCT MCHC less than 37 IF PATIENT IS NORMAL

Physiologic differences

Race, age, gender, geographic location

High platelet count is seen in:

Reactive to malignant conditions

Keratocyte (horn)

Red cells with one or several notches that look like horns on either end

Burr cells (echinocytes)

Result from blowing on the slides

Dacrocyte (Teardrop)

Round cell with a single elongated point

Basophilic Stippling

Round or irregularly shaped granules of variable number and size, composed of aggregates of ribosomes, stain blueish/black

Definitive Erythropoiesis

Self renewing hematopoietic stem cells (HSC's) produced

Howell-Jolly bodies

Small, round bodies composed of DNA, stains dark purple

Spherocytes

Smaller and conditioned by the spleen

Blue Tube Top

Sodium Citrate

Green Tube Top

Sodium Heparin -Rarely seen in hematology -Heparin stops/slows the action of thrombin

Spherocyte

Spherocytic red cells with dense hemoglobin, lack an area of central pallor

What's involved in bone making?

Spleen, liver, bone marrow

What tissues are considered secondary tissues?

Spleen, lymph nodes, MALT, GALT, BALT, SALT

Acanthocyte (spike)

Spurr cell, spicules of varying length irregularly distribute over the surface

QC (Quality Control)

Test to prove that the test is accurate

Reference Range

The numeric range of test values for which the general population consistently shows similar results 95% of the time

QC Range

The range in which a certain QC test result must fall to accept patient results

Codocyte (Bell/Target cell)

Thin, bell shaped that appears as a target with a central bulls eye

Leptocyte (thin)

Thin, flat cell with hemoglobin's, cup shaped

Platelets

Thrombocytes Clotting blood Necessary for homeostasis

What are the adult hemopoietic tissues?

Tissues involved in production or development: Bone marrow, lymph nodes, thymus, spleen, liver

Why do we need data ranges?

To make our data points mean something

The coulter uses what technology to determine the WBC populations?

VCS technology (Volume, Conductivity, Scatter)

4 systems involved in hemostasis:

Vascular, Platelets, Coagulation, Fibrinolytic

What parameters are measured directly on the counter?

WBC, RBC, Hgb

What does MCHC indicate?

Whether the RBC population is normochromic or hypochromic

3 Cell Types found in blood

erythrocytes, leukocytes, platelets

What are the five types of WBC's in blood?

neutrophils, lymphocytes, monocytes, eosinophils, basophils

Inclusions:

Basophilic Stippling, Cabot Rings, Howell-Jolly bodies

Chromia is

Color

Drepanocyte (Sickle)

Contain polymerized hemoglobin showing various shapes