KAIMOSI FRIENDS UNIVWERSITY

SCHOOL OF HEALTH SCIENCES

DEPARTMENT OF NURSING
COURSE TITTLE: HEMATOLOGY
COURSE CODE: NCN 216
LECTURER: MADAM SARAH MAKONG’O
TASK: GROUP 4 ASSIGNMENT
DATE OF SUBMISSION: 21ST NOVEMBER, 2022
GROUP PARTICIPANTS

GROUP MEMBER ADMISSION NO.

Churchill Sianzu HNR/0036/2022
Mitchelle Maiyo HNR/5003/2021
Brian Obwonyo HNR/0060/2021
Gail Kerubo HNR/0044/2022
Vincent Too HNR/0017/2022
Sharon Nyanchama HNR/0014/2022
Kevin Kiprop HNR/0019/2022
Carlson Kenyan HNR/0032/2022
Ryan Korir HNR/0022/2022
Rono Allan HNR/0046/2022
Lameck Mochama HNR/0048/2022

RED BLOOD CELL COUNT

It is a blood test that tells how many RBCs one has
RBCs contain HB which carries oxygen around
The amount of oxygen is delined to the body tissue depend on the number of RBCs one has and
how they work.
A normal RBC count:
 Men- 4.0 to 5.9*1012 /litre
 Women-3.8 to 5.2*1012/litre

FRAGILITY AND SUCKING TEST

Fragility test
The osmotic fragility test (OFT) is used to measure erythrocyte resistance to hemolysis while
being exposed to varying levels of dilution of a saline solution.
PRINCIPLE: Microcytic Red Blood Cells are resistant to lysis when exposed to hypotonic solution.
SAMPLE: Whole Blood in EDTA
REAGENTS: *0.36% buffered saline.
*Tyrode's solution
Procedure
1) 20 μl of whole blood collected in EDTA is pipetted into a glass test tube (100 × 10 mm)
containing 4 ml of 0.36% buffered saline solution.
2) Shake the tube and leave at room temperature for 30 minutes.
3) Shake again the tube and immediately hold the tube in front of a piece of paper with text
Interpretation : If the words on the paper are clearly visible through the tube, the test is
negative; whereas if the words are not clearly visible the test is positive (thalassaemia trait) due
to the turbidity of the solution.
Abnormal results indicate thalassemia and hereditary spherocytosis
Risks

 Excessive bleeding
 Fainting or feeling lightheaded
 Multiple punctures to locate veins
 Hematoma (blood buildup under the skin)
 Infection (a slight risk any time the skin is broken)

Sickling test
A sickle cell test is a simple blood test used to determine if you have sickle cell disease (SCD) or
sickle cell trait. People with SCD have red blood cells (RBCs) that are abnormally shaped. Sickle
cells are shaped like a crescent moon. Normal RBCs look like doughnuts.
Principle: Sodium metabisulphite reduces the oxygen tension inducing the typical sickle-shape
of red blood cells.
Sample: Fresh blood in any anticoagulant.
Reagents: 0.2 g of sodium metabisulphite in 10 ml of distilled water. Stir until dissolved.
Prepare fresh each time.
Procedure
 Mix 1 drop of blood with 1 drop of 2% sodium metabisulphite solution on a microscope
slide.
 Cover with a cover slip and seal the edge with wax/vaseline mixture or with nail varnish.
Allow to stand at room temperature for 1 to 4 hours.
 Examine under a microscope with the dry objective.

Results
In positive samples the typical sickle-shaped red blood cells will appear .Occasionally the
preparation may need to stand for up to 24 hours. In this case put the slides in a moist Petri
dish. False negative results may be obtained if the metabilsulphite has deteriorated or if the
cover slip is not sealed properly.
Risks
i. Excessive bleeding
ii. Fainting or feeling lightheaded
iii. Multiple punctures to locate veins
iv. Hematoma (blood buildup under the skin)
v. Infection (a slight risk any time the skin is broken)

Hb estimation
Hemoglobin is measured to detect anemia and its severity and to monitor an anemic patient’s
response to treatment. The test is also performed to check the hemoglobin level of a blood
donor prior to donating blood. Capillary blood or venous blood can be used. The hemoglobin
content of a solution may be estimated by several methods:
 by measurement of its color
 its power of combining with oxygen
 carbonmonoxide and by its iron content
Hemoglobin is measured photometrically or estimated using a visual comparative technique. In
photometric techniques the absorbance of hemoglobin in a blood sample is measured
electronically using a filter colorimeter or a direct read-out hemoglobin meter. When it is not
possible to measure hemoglobin accurately using a photometric technique a visual comparative
technique can help to detect anemia and assess its severity. Hemoglobin values can be
expressed in grams per liter (g/ l) or grams per deciliter (g/dl). Grams/liter is the recommended
way of expressing the mass concentration of hemoglobin.

CYANMETHEMOGLOBIN (HEMIGLOBINCYANIDE(HICN)) METHOD

This technique is ICSH recommended because stable hemiglobincyanide (HiCN) standards are
available to calibrate instruments. The technique is also used as a reference method against
which all other color comparison methods should be calibrated.
Principle of the test
Whole blood is diluted in a modified Drabkin’s solution which contains potassium ferricyanide
and potassium cyanide. The red cells are hemolyzed and the hemoglobin is oxidized by the
ferricyanide to methemoglobin (Hemiglobin, Hi). This is converted by the cyanide to stable
hemiglobin cyanide (HiCN). Absorbance of the HiCN solution is read in a spectrophotometer at
wavelength 540nm or in a filter colorimeter using a yellow-green filter. The absorbance
obtained is compared with that of a reference HiCN standard solution. Hemoglobin values are
obtained from tables prepared from a calibration graph or if using a direct read-out hemoglobin
meter, for the digital display.
Test method

1. Measure carefully 20µl (0.02ml) of capillary blood or well-mixed venous blood and dispense
it into 3.98ml Drabkin’s neutral diluting fluid.
2. Stopper the tube, mix, and leave the diluted blood at room temperature, protected from
sunlight, for 4-5 minutes. This time is adequate for conversion of hemoglobin to HiCN when
using a neutral (pH 7.0-7.4) Drabkin’s reagent. Up to 20 minutes is required when using an
alkaline Drabkin’s reagent.
3. Place a yellow-green filter in the colorimeter or set the wavelength at 540nm
4. Zero the colorimeter with the ammonia water diluting fluid. Read the absorbance of the
patient’s sample.
5. Using the table prepared form the calibration graph, read off the patient’s hemoglobin value.

Oxyhemoglobin Method
 This is a reliable and inexpensive method of measuring hemoglobin but there is no stable
oxyhemoglobin (HbO2) reference standard solution available for the direct calibration of the
HbO2 technique. Preparation of a calibration graph for use with a filter colorimeter, requires
the use of a secondary blood standard, i.e. a whole blood or hemolysate of known hemoglobin
value (between 140-160g/l) that has been previously analyzed by the HiCN method or other
well calibrated technique.
Principle of test
Blood is diluted in a weak ammonia solution. This lyzes the red cells. The absorbance of the
solution is measured as oxyhemoglobin in a filter colorimeter using a yellow-green filter or at
wavelength 540nm. Hemoglobin values are obtained from tables prepared from a calibration
graph. Methemoglobin and carboxyhemoglobin are not accurately detected but these are
normally present only in trace amounts and are not oxygen-carrying forms of hemoglobin

Test method
1. Measure carefully 20µl of capillary or well-mixed venous blood and dispense into 3.98ml
(4ml) ammonia water diluting fluid
2. Stopper the tube and mix well.
3. Place a yellow-green filter in the colorimeter or set the wavelength at 540nm.
4. Zero the colorimeter with the ammonia water diluting fluid. Read the absorbance of the
patient’s sample.
5. Using the table prepared from the calibration graph, read off the patient’s hemoglobin value

Acid Hematin Method (Sahli-Hellige)

This visual comparative method of estimating hemoglobin although still used in some health
centers and hospitals is not recommended because of its unacceptable imprecision and
inaccuracy. Most of the problems associated with the Sahli method are due to the instability of
acid hematin, fading of the color glass standard and difficulty in matching it to the acid hematin
solution. Conversion to acid hematin is slow. HbF is not converted to acid hematin and
therefore the Sahli method is not suitable for measuring hemoglobin levels in infants up to 3
months.
Principle
Hemoglobin in a sample of blood is converted to a brown colored acid hematin by treatment
with 0.1 N HCl and after allowing the diluted sample to stand for 5 minute to ensure complete
conversion to acid hematin it is diluted with distilled water until its color match as with the
color of an artificial standard (tinted glass).
Materials
Sahli hemoglobinometer
Sahli pipette
Stirring glass rod
Dropping pipette
Absorbent cotton 0.1N HCl
Test method
 1. Fill the graduated tube to the ''20'' mark of the red graduation or to the 3g/dl mark of the
yellow graduation with 0.1N HCl.
2. Draw venous or capillary blood to the 0.02ml mark of the Sahli pipette. Do not allow air
bubbles to enter. Do not take the first drop of blood from the finger.
3. Wipe the outside of the pipette with absorbent paper. Check that the blood is still on the
mark.
4. Blow the blood from the pipette into the graduated pipette into the graduated tube of the
acid solution. Rinse the pipette by drawing and blowing out the acid solution 3 times. The
mixture of the blood and acid gives a brownish color. Allow to stand for 5 minutes.
5. Place the graduated tube in the hemoglobinometer stand facing a window.Compare the
color of the tube containing diluted blood with the color of the reference tube. If the color of
the diluted sample is darker than that of the reference, continue to dilute by adding 0.1N HCl or
distilled water drop by drop. Stir with the glass rod after adding each drop. Remove the rod and
compare the colors of the two tubes. Stop when the colors match.
6. Note the mark reached. Depending on the type of hemoglobinometer, this gives the
hemoglobin concentration either in g/dl or as a percentage of ''normal''. To convert
percentages to g/dl, multiply the reading by 0.146.
Interpretation of hemoglobin test results
Normal hemoglobin levels vary according to age and gender, and the altitude at which a person
lives. Normal hemoglobin reference range:
Children at birth 135-195 g/l
children 2 y – 5 y 110-140 g/l
Children 6 y – 12 y 115-155 g/l
Adult men 130-180 g/l
Adult women 120-150 g/l

Erythrocyte sedimentation rate (ESR)

Erythrocyte sedimentation rate (ESR), is a blood test that can show inflammatory activity in the
body. Many health problems can cause a ESR test result to be outside the standard range.
It is often used with other tests to help your health care team diagnose or check the progress of
an inflammatory disease.
When blood is placed in a tall, thin tube, red blood cells, called erythrocytes, gradually settle to
the bottom. Inflammation can cause the cells to clump together.
Because these clumps are heavier than individual cells, they settle to the bottom more quickly.
ESR test measures the distance red blood cells fall in a test tube in one hour.
The farther the red blood cells have fallen, the greater the inflammatory response of your
immune system.
Factors Affecting ESR
 Technical factors: short ESR tubes, low room temperature, delay in test performance (>2
hours), clotted blood sample, excess anticoagulant, bubbles in tube.
 Chronic inflammatory disease (collagen and vascular diseases) increases ESR.
 Polycythemia decreases ESR.
 Anemia increases ESR because the change in erythrocyte-plasma ratio favors rouleaux
formation.
 Increase level of fibrinogen increases ESR.

WHITE BLOOD CELL COUNT

WBC count is a blood test that measures the number of white blood cells in your blood.
White blood cells are part of the immune system and help your body fight off infections and
other diseases.
The normal WBC count ranges between 4,000 and 11,000 per microliter of blood, but it may
vary for men, women, and children.
A low WBC count could mean your body may not be able to fight infection the way it should

Platelet count
A platelet count measures the average platelet level in a person’s blood. High or low platelet
levels can increase the risk of clotting or excessive bleeding.
Platelets are fragments of larger cells made in the bone marrow called megakaryocytes. These
fragments are crucial to wound healing.
Abnormal platelet levels can lead to various health complications. This article discusses the
process of a platelet blood test and what the results mean.
The mean platelet count blood test is typically part of a complete blood count (CBC) test. A CBC
reveals important information about the number of different blood cells in the body.
A platelet count test reveals the average number of platelets a person has per microliter of
blood.
The test involves drawing blood from a vein in the arm or hand.
Obtaining a blood sample from a vein takes a few minutes and generally causes only minimal
discomfort. Occasionally, some people may feel queasy or light-headed while the blood is
drawn or shortly after. Taking slow deep breaths is usually enough to calm these feelings. Some
people may develop a small mark or bruise.
Typically, doctors will send these samples to a lab for assessment and relay results to the
patient in due course.
Normal platelet levels
A person’s platelet levels can change with age, and certain medical conditions can also affect
them.
A platelet count that is too low or too high can lead to health complications. A low platelet
count is known as thrombocytopenia, while a high platelet count is known as thrombocytosis.
Tests measure average platelet levels per microliter (mcL) of blood. Below are guideline platelet
levels.
Result Platelet count

high platelet level (thrombocytosis) more than 450,000

normal platelet level 150,000–450,000

low platelet level (thrombocytopenia) less than 150,000

What does it mean when your platelet count is low?
A low platelet count can make it difficult for the blood to clot, putting a person at risk of
excessive bleeding. Low platelet counts can be due to an inherited medical condition or an
acquired medical condition. In some cases, the cause may be unknown.
Risks and complications
 Bleeding spontaneously. This is a medical emergency, and people who experience
spontaneous bleeding may require a blood, or platelet, transfusion.
 Low platelet count increases the risk of death in people who experience a traumatic injury.
High platelet count
A high platelet count can occur when something causes the bone marrow to make too many
platelets. When the reason is unknown, it is called primary or essential thrombocytosis.
When excess platelets are due to an infection or other condition, it is called secondary
thrombocytosis.
Risks and complications
A person’s blood clots more quickly when they have too many platelets. Clotting is a natural
protection against bleeding. The body produces more platelets during and following an injury.
However, because platelets cause blood clotting, they can also cause dangerous blood clots in
the arms or legs. The blood clot may break off or travel to another area of the body.
The risk of a blood clot is higher in people confined to bed by illness or who cannot move their
limbs.
Someone who has a high platelet count because of a recent injury but who must remain in bed
may need monitoring to reduce the risk of blood clots as a result.
Causes
Several factors can cause a person’s platelet levels to change. These include acute and chronic
medical conditions and age.
Causes of high platelet counts
Some temporary conditions can cause a higher-than-normal platelet count. A doctor may order
a retest a few days or weeks later if this happens. Some common reasons for high platelet
levels include:
 recovering from a recent injury
 recovering from blood loss after surgery
 recovering from excessive drinking or vitamin B12 deficiency
 intense physical activity or exertion, such as from running a marathon
 using birth control pills
If a person’s platelet count remains high, chronic medical conditions may be responsible. These
may include:
 cancer
 iron deficiency anemia
 inflammatory disorders
 infections, such as tuberculosis
 splenectomy, or a spleen removal
Common causes of low platelet volume include:
 viruses such as mononucleosis, HIV, AIDS, measles, and hepatitis
 medications such as aspirin, H2-blockers, and quinidine
 cancer that spreads to the bone marrow
 aplastic anemia
 bacterial infection such as sepsis
 autoimmune diseases such as lupus and Crohn’s disease
 chemotherapy harms existing tissue and cancer cells, making it difficult for the body to
produce platelets.
 liver cirrhosis
 chronic bleeding
 Platelet count tends to decline with age. A platelet count that is lower than it once was or on
the lower end of normal may not be a cause for concern in an older adult — especially if
there are no other symptoms.