Week 1: Handout

Write a short statement about each of the following tests and mark which would be available
in rural and remote laboratories and which is part of a standard full blood count?

Hb:

 Haemoglobin level in blood

 Male 130 – 180 g/L

 Female 115 – 165 g/L

 Neonate (Full term) 135 – 195 g/L

 Part of standard

Hb electrophoresis:

 In hemoglobin electrophoresis, hemoglobins under the influence of an electrical field

exhibit varying mobilities, which allows differentiation of haemoglobin.

Hct:

Hematocrit is the ratio of the volume of RBCs to the volume of whole blood and is
determined by transferring blood to a graduated plastic tube, centrifuging, measuring the
column of RBCs, and dividing by the total length of RBCs plus plasma. The normal ratio
approaches 50%. Hematocrit is also called packed cell volume (PCV), the packed cells
referring to RBCs.

MCV:

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MCH

MCHC

Reticulocyte count

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ESR

Red cell count

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RDW

Platelet count

The purpose of platelets

“Platelets are the cells that circulate within our blood and bind together when they recognize
damaged blood vessels,” says Dr. Marlene Williams, Assistant Professor of Medicine and
CICU Director for Johns Hopkins Bayview Medical Center. “When you get a cut, for
example, the platelets bind to the site of the damaged vessel, thereby causing a blood clot.
There’s an evolutionary reason why they’re there. It’s to stop us from bleeding.”

What makes platelets change their shape?

Platelets, the smallest of our blood cells, can only be seen under a microscope. They’re
literally shaped like small plates in their non-active form. A blood vessel will send out a
signal when it becomes damaged. When platelets receive that signal, they’ll respond by
traveling to the area and transforming into their “active” formation. To make contact with the
broken blood vessel, platelets grow long tentacles and then resemble a spider or an octopus.

What is a healthy platelet count?

A normal platelet count ranges from 150,000 to 450,000 platelets per microliter of blood.
Having more than 450,000 platelets is a condition called thrombocytosis; having less than
150,000 is known as thrombocytopenia. You get your platelet number from a routine blood
test called a complete blood count (CBC).

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White cell count

White cell differential

Reference Range

Differential blood count gives relative percentage of each type of white blood cell and also
helps reveal abnormal white blood cell populations (eg, blasts, immature granulocytes, or
circulating lymphoma cells in the peripheral blood).

CD markers – cluster of differentiation

Cell surface membrane receptors or markers used to characterize cells by their functions. CD
profiles are used in flow cytometry to identify cell types. CDs are used in hematology to
identify cell clones associated with lymphatic and myelogenous leukemias and lymphomas.

PT

The prothrombin time (PT)—along with its derived measures of prothrombin ratio (PR) and
international normalized ratio (INR)—are assays evaluating the extrinsic pathway of
coagulation. This test is also called "ProTime INR" and "PT/INR". They are used to
determine the clotting tendency of blood, in the measure of warfarin dosage, liver damage,
and vitamin K status. PT measures factors I (Fibrinogen), II (Prothrombin), V (Proaccelerin),
VII (Proconvertin), and X (Stuart–Prower Factor). It is used in conjunction with the activated
partial thromboplastin time (aPTT) which measures the intrinsic pathway and common
pathway.

APTT

Partial thromboplastin time (PTT) and activated partial thromboplastin time (aPTT) are used
to test for the same functions; however, in aPTT, an activator is added that speeds up the
clotting time and results in a narrower reference range. The aPTT is considered a more
sensitive version of the PTT and is used to monitor the patient’s response to heparin therapy.

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Fibrinogen

This testing is used to evaluate fibrinogen, a protein that is essential for blood clot formation.
When there is an injury and bleeding occurs, the body forms a blood clot through a series of
steps. In one of the last steps, soluble fibrinogen is converted into insoluble fibrin threads that
crosslink together to form a net that stabilizes and adheres at the injury site until the area has
healed.

D-dimers
One of the fibrin degradation products. D-dimer is composed of two fibrin D fragments
covalently joined by factor XIII. The D-dimer assay is used to rule out venous
thromboembolic disease and disseminated intravascular coagulation, and may be used to
monitor the efficacy and length of warfarin therapy.

Coagulation factor assays

Bone marrow analysis

Bone marrow examination refers to the pathologic analysis of samples of bone marrow
obtained by bone marrow biopsy (often called a trephine biopsy) and bone marrow aspiration.
Bone marrow examination is used in the diagnosis of a number of conditions, including
leukemia, multiple myeloma, lymphoma, anemia, and pancytopenia. The bone marrow
produces the cellular elements of the blood, including platelets, red blood cells and white
blood cells. While much information can be gleaned by testing the blood itself (drawn from a
vein by phlebotomy), it is sometimes necessary to examine the source of the blood cells in

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the bone marrow to obtain more information on hematopoiesis; this is the role of bone
marrow aspiration and biopsy.

Cytogenetics

Sources of blood for analysis

 Arterial

 Venous

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 Capillary

 Bone marrow

What factors can reduce the quality of the blood sample?






Which stains are routinely used in haematology?

– May-Grünwald Giemsa stain

• >30 minutes; Great morphology
– Wright’s stain
• 10 min; good morphology
– Leishman’s stain
• 10 min; good morphology
– Field’s stain (diff-quick)
• 15 seconds; not good morphology

What are the functions of blood?

Blood plays an important role in regulating the body's systems and maintaining homeostasis.
It performs many functions within the body, including:

 Supplying oxygen to tissues (bound to hemoglobin, which is carried in red cells)

 Supplying nutrients such as glucose, amino acids, and fatty acids either dissolved in the blood
or bound to plasma proteins (e.g., blood lipids)
 Removing waste such as carbon dioxide, urea, and lactic acid
 Immunological functions, including circulation of white blood cells and detection of foreign
material by antibodies

 Coagulation, which is one part of the body's self-repair mechanism (blood clotting by the
platelets after an open wound in order to stop bleeding)

 Messenger functions, including the transport of hormones and the signaling of tissue damage

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 Regulating body pH

 Regulating core body temperature

 Hydraulic functions, including the regulation of the colloidal osmotic pressure of blood

What is blood comprised of?

Your blood is made up of liquid and solids. The liquid part, called plasma, is made of water,
salts, and protein. Over half of your blood is plasma. The solid part of your blood contains
red blood cells, white blood cells, and platelets.

Red blood cells (RBC) deliver oxygen from your lungs to your tissues and organs. White
blood cells (WBC) fight infection and are part of your immune system. Platelets help blood to
clot when you have a cut or wound. Bone marrow, the spongy material inside your bones,
makes new blood cells. Blood cells constantly die and your body makes new ones. Red blood
cells live about 120 days, and platelets live about 6 days. Some white blood cells live less
than a day, but others live much longer.

There are four blood types: A, B, AB, or O. Also, blood is either Rh-positive or Rh-negative.
So if you have type A blood, it's either A positive or A negative. Which type you are is
important if you need a blood transfusion. And your Rh factor could be important if you
become pregnant - an incompatibility between your type and the baby's could create
problems.

Where is blood made?

Haematopoiesis

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Why do red cells stain red and Neutrophils stain purple?

 Methylene blue
Stains acidic elements (eg: nuclei)
 Eosin Y
Stains alkali and neutral elements (eg: red cells)

Match these cell pictures as seen down the microscope with the cell type
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