General Hematology  

Laboratory Report  
Based On: Red Blood Cell Count 

Name of Student : Romanda Greene

USI # 1019820
Date: 22-10-2018
Title: Red Blood Cell Count ( Manual Method)

Principle: ​Anticoagulated whole blood is centrifuged, and the volume occupied by the erythrocytes are
merely expressed as a percentage of the total volume.[1,2]The centrifuge, through centrifugal forces,
separates formed elements from liquid plasma.[3] The microhematocrit centrifuge quickly attains
speeds of 11,000 rpm and RCFs of up to 15,000 g to spin the heparinized microcapillary tube
samples.[3] As a result of it’s density, erythrocytes settles to the bottom, leukocytes and platelets settles
at the middle forming the buffy coat and liquid plasma at the top layer of the packed cell column.[4]
Haematocrit is determined by the volume of plasma as compared to the volume of erythrocytes.[4]

Material and Reagents: as per lab handout

Method: as per lab handout
Clinical Significance: The microhematocrit is typically a routinely performed test -ordered as a part of
the complete blood count (CBC) - that adequately provides the practitioner with an modest estimate of the
patient’s red blood cell volume [5,6,]. Thus, the blood’s oxygen carrying capacity.[5,6,] Generally,
plasma contributes 55% and formed elements 45% ( with leukocytes and platelets contributing 1%) of the
total volume of whole blood. [4]
Since, a hematocrit is often performed as part of a complete blood count (CBC) , results from other
components, like RBC count, hemoglobin, reticulocyte count, and/or red blood cell indices, are taken into
consideration.[6] Age, sex, and race remain other factors to be considered. Habitually, the hematocrit
mirrors the results of the RBC count and hemoglobin.[6]

Abnormal High [A hematocrit (Hct) value that is above the maximum limit of the standard range]
demonstrates an absolute increase in the number of erythrocytes and a decline in the plasma volume.[7]
Elevated packed cell volume indicates Polycythemia or Dehydration,[5-8] thus, an increase in the
viscosity of the blood, [8]which makes it more difficult for the heart to circulate the blood.[8]

Some probable causes of a high hematocrit include:

Chronic Obstructive Pulmonary Disease (COPD) - this includes Chronic Bronchitis, Emphysema and
Refractory (non-reversible) Asthma - is typically a chronic inflammatory lung disease that causes
obstructed airflow from the lungs.[9] The patient is incompetent to breathe and absorb sufficient oxygen,
resulting in an increase in erythropoietin[5]: stimulating red blood cell production in the bone marrow to
adequately compensate for Hypoxia. [5]

Ventricular septal defect (VSD) - A condition in which there is a defect (hole) in the septum that separates
the heart's lower chambers (ventricles) and allows blood to pass from the left to the right side of the heart.
[10] The oxygen-rich blood then gets pumped back to the lungs instead of out to the body.[10]Leading to
reduced oxygen levels in the blood. The body tries compensating by producing more red blood cells.[5]

Polycythemia Vera a myeloproliferative blood condition that occur when the bone marrow produces
excess quantities of erythrocytes (Hyperplasia).[5]

Signs and symptoms of polycythemia typically include: Disturbed ( double/Blurred) vision, Dizziness,
Headache, Flushing, Enlarged spleen, Tiredness, Excessive sweating and Itching [5-8].

Abnormal Low: [A hematocrit (Hct) value that is below the lower limit of the normal range] reflect a
decrease in the number of erythrocytes and an increase in the plasma volume.[7] A curtailing in the
packed cell volume indicates Anemia[5-8], a state where there is a depletion in the number of erythrocytes
or their oxygen carrying capacity is insufficient[5].

Some causes of a low hematocrit include:

Kidney Failure In cases like Acute glomerulonephritis, Acute Tubular Necrosis, Interstitial Cystitis and
Nephropathic Cystinosis leads to a depletion of erythropoietin,[11] a hormone produced by the
functioning kidneys that stimulates RBC production by the bone marrow.[5]

Menorrhagia - Heavy menstrual bleeding can undoubtedly cause a significant loss of iron-rich blood,
resulting in iron deficiency anemia.[9] Iron is an essential nutrient that is needed in small quantities to
help form normal red blood cells and is a critical part of hemoglobin. [10-11] When the level of iron is
insufficient iron stores are depleted, the bone marrow is not able to increase production of new red blood
cells to replace those lost.[5,9, 10,11]
Aplastic Anemia-a blood disorder in which the body's bone marrow does not produce enough new blood
cells due to acquire or inherited damage to the bone-marrow stem cells.[5] This embodies radiation and
chemotherapy, medications, such as Chloramphenicol, infectious diseases, such as Hepatitis, Epstein-Barr
virus and HIV, autoimmune disorders, like Lupus and Rheumatoid Arthritis.[10-11]

Signs and symptoms of anemia prominently include: Weakness or fatigue, Lack of energy, Fainting,
Paleness (pallor), Shortness of breath, Cold feet and Hands and Chest Pain. [5]

A hematocrit maybe ordered in cases of severe dehydration, like extreme thirst, dry mouth or mucous
membranes, burns, diarrhoea and lack of sweating or urination and during pregnancy. [5]

This test may be performed several times or on a frequent basis when someone has been diagnosed with
ongoing bleeding problems, anemia, or polycythemia to determine periodic the effectiveness of
treatment.[5] It may also be ordered routinely for people undergoing treatment for cancer known to affect
the bone marrow.[5]

Limitations :

A number of factors can affect the outcome of a hematocrit test and yield inaccurate or misleading
results, including:

❏ Losing blood from whatever cause will cause hematocrit levels to decrease. [5-7]
❏ People living in high altitudes have a high hematocrit value.[5-7]
❏ Normally, hematocrit value slightly decreases in the physiologic Hydremia of pregnancy.
[5-7]
❏ The normal hematocrit value for newborns is higher because there is many macrocytic red
cells. [5]
❏ There is tendency towards lower hematocrit value in men and women older than 60 years of
age , corresponding to lower red blood cell values in this age group.[5]
❏ Dehydration from any cause falsely raises the hematocrit value because of fluid loss,
which causes a decrease in plasma volume.[5-7]
❏ If blood is drawn from a capillary puncture and a microhematocrit is done, values are
slightly higher. [5]
❏ Difficult venipuncture or skin puncture may introduce interstitial fluid to the sample,
causing a falsely decreased Hematocrit value. [7]
 ❏ When obtaining blood specimen, leaving the tourniquet on the arm too long causes
hemoconcentration, which falsely increase hematocrit values. [7]
❏ Inadequate centrifugation of the capillary tube or allowing the the tube to stand too long
after the centrifuge has stopped increases the hematocrit value. [5-7]
❏ Hemolysis also causes a false decrease in hematocrit values.[5-7]

Sources of Error:

Erroneous results can be caused based on the following:

❏ Improper sealing of capillary tubes causes a decrease hematocrit values to decrease due to loss
of erythrocytes. [7]

❏ Underfilled original sample due to excessive anticoagulant causes erythrocytes to shrink ,

therefore, the hematocrit value decreases. [7]

❏ Not having the ability to interpret the different layers of the packed cell volume column:thus,
resulting in the inclusion of the buffy coat in reading the packed cell column.[7,12]

❏ Insufficient mixing of blood before obtaining the hematocrit value. [7,12]

❏ Allowing the capillary tube to stand and not allowing it to be sufficiently centrifuged resulting
in improper packing. Therefore, there is an increase in the packed cell volume
(hematocrit)[7,12]

❏ Improper use of the HCT reader may increase or decrease the hematocrit value, due to
parallax. [7,12]

Normal Range
Children 1- 10 years 31-41% [7]
Adults
Men 40-54 % [7]
Women 38-47% [7]
Critical Values : < 15% or > 60% [7]
Quality Control
 ❏ The centrifuge should be checked regularly for timer accuracy and reproducibility with a
stopwatch and Speed should be check with a properly calibrated tachometer.[12]

❏ Centrifuges should be check for the minimal time for optimal maximal packing of cells. [12]

❏ Centrifuges brushes should be checked regularly and replaced when the brushes are less than
⅟ 2 their original size. [12]

❏ Commercially available whole blood ( control) with know value can be used to check the
accuracy of normal and abnormal levels and run on a daily basis. [12]

❏ All test should be done in duplicate to eliminate errors caused by leakage. [12]

❏ Verify that results obtained is within range for control used. The results done in duplicate
hematocrit should agree within ± 2% . if results obtained are not within its tolerance limit, the
entire procedure must be repeated. [12]

❏ Buffy coat should not be included. [12]

Results: After 1 -hour of leaving the capillary tube to stand and allowing the packed cell volume
column to form (i.e the red blood cell settles at the bottom of the capillary tube and liquid plasma at
the top layer) , the packed cell volume (hematocrit value) was 75%.

Plasma seems to have a ​brown or orange tinge and the buffy coat was not visible.

Conclusion: Polycythemic state is indicated when ​hematocrit values that are elevated and the plasma
volume does not increase. An anemic state is considered when hematocrit values decreases and the
plasma volume increase.

References:

1. International Committee for Standardization in Hematology. Selected method for determination of

Packed Cell volume by the microhematocrit method. 2nd Edition. Villanova, PA. NCCLS.1993
pg.72-75

2.National Committee for Clinical Laboratory Standards Procedures for determining packed cell
volume. In advances in Hematologic Methods:The blood count. Fl. CRC Press.1952 pg. 30

3. World Health Organization. ECRI Institute’s Health Care.2012

http://www.who.int/medical-devices
4. Bett JG, Johnson JE, Wise JA et.al. Anatomy and Physiology. 4th edition. Heyden JR. Openstax
College. 2013.pg. 784-786. ​http://cnx.org/contents/Fptklzmh@8.25​.

5. The American Association for Clinical Chemistry (AACC). Hematocrit. Schaeffer,K. AACC
Publishers.2017. ​http://labtestonline.org/tests/hematocrit​.

6.Pruthi, RK. Hemoglobin Test. Pruthi, S. MayoClinic. 2011

https://www.mayoclinic.org/tests-procedures/hemoglobin-test/about/pac-20385075

7. Daniels R. Delmar’s Guide to Laboratory and Diagnostic Tests.2nd Edition. New York, New
York. Delmar Cengage Learning.2010 pg. 394-395

8. Sampson, S. All you need to know about thick blood. Whiteman H, Newman T, Martin VH.
Medical News Today.2017. ​https://www.medicalnewstoday.com/articles/319842.php

9.Suzanne CL,Bonnie F, Paula, M. Chronic Obstructive Pulmonary Disease (COPD).Vol.17. Wilson

K, ZuWallack, R. The American Thoracic Society. 2005 .pg 3-4
https://www.thoracic.org/patients/patient-resources/resources/copd-intro.pdf

10.Center for Disease Control (CDC). Facts about Ventricular Septal Defect. Atlanta, GA. US
Department for Human Health and Services. 2018.

https://www.cdc.gov/ncbddd/heartdefects/ventricularseptaldefect.html

11. Brady H.R, Brenner B.M, Lieberthal W. Acute renal failure. In: The Kidney 5th Ed.; Brenner,
B.M., Ed.; Philadelphia, Saunders, 1996; pg.1200–1252.

12. Frances TF, Marshall BD. A manual of Laboratory Diagnostic Test. 7th Edition. Lippincott
Williams and Wilkins.2004 pg.132-133