Determination of Arterial Blood Pressure by Auscultatory Method
Discussion
Blood pressure is the force exerted by circulating blood against the walls of blood vessels. It
consists of two components: systolic pressure (normally around 120 mmHg), recorded during
ventricular contraction, and diastolic pressure (around 80 mmHg), recorded during ventricular
relaxation. In this experiment, blood pressure was measured using a sphygmomanometer and
stethoscope by both palpatory and auscultatory methods.
The cuff was wrapped around the left arm at heart level, ensuring proper fitting for accuracy,
since the heart lies slightly toward the left side of the thorax. The palpatory method helped
estimate the systolic pressure, after which the cuff was inflated about 20 mmHg above this value
to perform the auscultatory method. The appearance of the first Korotkoff sound indicated
systolic pressure, while the disappearance of the sound indicated diastolic pressure.
Our results were within the normal range, indicating normal cardiac output and peripheral
resistance. Minor variations among subjects could be due to anxiety, posture, cuff tightness, or
recent physical activity. The auscultatory method provided more precise readings compared to
the palpatory method, which only gives systolic pressure. Direct measurement using arterial
cannulation is considered the gold standard but is reserved for critical care due to its invasive
nature.
Determination of bleeding time of own blood
Discussion
The bleeding time test assesses the efficiency of platelets and the integrity of capillary
endothelium, reflecting the body’s primary hemostatic response. In this experiment, the bleeding
time was approximately 3 minutes, which falls within the normal range of 2–5 minutes. This
finding indicates normal platelet function and vascular integrity. A prolonged bleeding time may
suggest platelet deficiency, qualitative platelet dysfunction, vascular disorders, or the use of
antiplatelet drugs such as aspirin. This practical reinforced the understanding of primary
hemostasis, emphasizing the coordinated role of vascular constriction and platelet plug formation
in the prevention of blood loss following injury.
Determination of Pulse Rate
Discussion
The pulse rate represents the number of heart beats per minute and serves as an important
indicator of cardiac activity and circulatory efficiency. In this practical, the pulse was palpated at
the radial artery, and the observed pulse rate was within the normal range of 60–100 beats per
minute, indicating a normal cardiac rhythm and adequate peripheral perfusion.
Variations in pulse rate may occur due to factors such as physical activity, emotional stress,
temperature, or underlying conditions. A rate above 100 beats per minute (tachycardia) may be
seen in fever, hyperthyroidism, or anxiety, whereas a rate below 60 beats per minute
(bradycardia) may occur in athletes, hypothyroidism, or heart block. In addition to rate, other
characteristics such as rhythm, volume, tension, and equality were assessed to evaluate cardiac
and vascular function.
This practical enhanced our understanding of the relationship between heart rate, cardiac output,
and vascular health, emphasizing the clinical value of pulse examination in assessing
cardiovascular status.
Determination of Heart Rate Before and After Exercise
Discussion
Heart rate is the number of times the heart beats per minute and reflects the activity of the
cardiovascular and autonomic nervous systems. In this experiment, heart rate was recorded
before and after physical exercise to study the effect of muscular activity on cardiac function.
The observed heart rate increased immediately after exercise, indicating activation of the
sympathetic nervous system and inhibition of the parasympathetic tone. This response enhances
cardiac output and ensures adequate oxygen and nutrient delivery to active muscles. The rate
gradually returns to baseline during the recovery phase due to parasympathetic reactivation and
metabolic normalization.
This experiment demonstrates how heart rate regulation maintains homeostasis during increased
metabolic demand, and highlights the importance of autonomic control in adapting cardiac
activity to physiological stress such as exercise.
Determination of Clotting Time
Discussion
Clotting time is the duration required for blood to coagulate after it has been shed from the body,
reflecting the efficiency of the intrinsic pathway of coagulation and the activity of plasma
clotting factors. In this experiment, the clotting time was found to be within the normal range of
5–11 minutes, indicating normal function of coagulation factors and calcium ions.
A prolonged clotting time may occur in conditions such as hemophilia, liver disease, vitamin K
deficiency, or during anticoagulant therapy (e.g., heparin). A shortened clotting time is rare and
usually clinically insignificant.
This practical reinforced understanding of secondary hemostasis, emphasizing how plasma
proteins, platelets, and calcium ions interact to form a stable fibrin clot that prevents excessive
blood loss following vascular injury.
Determination of Clotting Time
Discussion
The electrocardiogram (ECG) records the electrical activity of the heart and provides valuable
information about its rhythm, conduction, and overall function. It represents the depolarization
and repolarization of cardiac muscle during each cardiac cycle. A normal ECG tracing consists
of a P wave (atrial depolarization), QRS complex (ventricular depolarization), and T wave
(ventricular repolarization).
In this practical, a normal sinus rhythm was observed, characterized by regular P waves
preceding each QRS complex and a heart rate within the normal range of 60–100 beats per
minute. The intervals and segments were within normal limits, indicating normal conduction
through the sinoatrial (SA) node, atrioventricular (AV) node, and ventricles.
Abnormalities in the ECG may indicate arrhythmias, myocardial ischemia, conduction blocks, or
electrolyte imbalances. This experiment helped us understand how electrical changes in the heart
correspond to mechanical events of the cardiac cycle and how ECG serves as a crucial diagnostic
tool in cardiovascular assessment.
Determination of Leukocyte Count of Blood
Discussion
The leukocyte count measures the number of white blood cells (WBCs) present in the blood and
serves as an indicator of the body’s immune status. In this experiment, the total leukocyte count
was found within the normal range of 4,000–11,000 cells/mm³ of blood, suggesting a normal
immune response.
An increase in leukocyte count (leukocytosis) may occur in infections, inflammation, stress, or
leukemia, whereas a decrease (leukopenia) may be seen in viral infections, bone marrow
suppression, or due to certain drugs such as cytotoxic agents.
This practical enhanced our understanding of the role of leukocytes in defense mechanisms and
how variations in their count can aid in the diagnosis and monitoring of disease conditions
affecting the hematopoietic and immune systems.
Estimation of Haemoglobin by Sahli’s Method
Discussion
Haemoglobin is the iron-containing pigment of red blood cells responsible for the transport of
oxygen from the lungs to the tissues and carbon dioxide from the tissues back to the lungs.
Estimating its concentration provides important information about the oxygen-carrying capacity
of blood and is routinely used to detect anaemia or polycythaemia.
In this experiment, haemoglobin concentration was estimated by Sahli’s (acid haematin) method.
The principle is based on the conversion of haemoglobin to brown acid haematin when blood is
mixed with hydrochloric acid; the resulting colour is then diluted with distilled water until it
matches the standard comparator. The intensity of colour corresponds to the haemoglobin
concentration, which is read directly in grams per decilitre (g/dL).
The observed haemoglobin values in this practical were within the normal physiological range—
approximately 13–17 g/dL in males and 12–15 g/dL in females, indicating adequate
erythropoietic activity and normal iron status among the participants. Minor variations between
individuals may arise due to differences in sex, altitude of residence, nutritional iron intake, or
hydration status. A decreased haemoglobin value suggests anaemia, which may result from iron
deficiency, chronic disease, blood loss, or bone-marrow disorders. Conversely, increased levels
can occur in dehydration, polycythaemia, or people living at high altitude due to chronic
hypoxia.
Although Sahli’s method is simple and inexpensive, its accuracy is limited by visual colour
matching and potential operator error. More precise techniques, such as the cyanmethemoglobin
method or automated haematology analysers, are preferred in clinical laboratories.
This experiment reinforced understanding of the functional importance of haemoglobin and the
clinical significance of its estimation in assessing oxygen transport and diagnosing
haematological disorders.
To study hemocytometer (improved neubaur’s chamber/ Levy’s chamber)
Discussion
A hemocytometer is a precision instrument used for the manual counting of blood cells,
originally designed for determining the number of red and white blood cells per cubic millimeter
of blood. The improved Neubauer’s or Levy’s chamber consists of a thick glass slide with a
finely ruled grid etched into its central platform and a defined depth of 0.1 mm when covered
with a special cover slip. The ruled area is divided into smaller squares that facilitate systematic
counting of cells under the microscope.
In this practical, the structure, design, and principle of the hemocytometer were studied to
understand its application in hematological estimations. The accurate use of the chamber
depends on several factors, including proper mixing of blood with the diluting fluid, correct
filling of the chamber by capillary action without air bubbles, and allowing cells to settle evenly
before counting. Understanding the grid layout—especially the central RBC counting area and
the four large corner squares for WBC counting—is essential for reliable calculations.
The study highlighted that accuracy in cell counting relies on maintaining standard dilution,
uniform distribution of cells, and correct mathematical computation of the final cell
concentration using the chamber’s known dimensions and dilution factors. Errors may occur due
to improper pipetting, overfilling or underfilling the chamber, uneven cell distribution, or failure
to use the correct depth of focus during microscopic examination.
This exercise enhanced understanding of the principle of quantitative microscopy and the
importance of standardization in hematological procedures. Although automated cell counters
are widely used today for greater accuracy and speed, the hemocytometer remains a fundamental
laboratory tool, providing students with essential insight into the manual techniques and
calculations underlying modern hematology.
Calculation of blood indices
Discussion
Blood indices are calculated parameters that provide valuable information about the size,
hemoglobin content, and concentration of red blood cells (RBCs). These indices include the
Mean Corpuscular Volume (MCV), Mean Corpuscular Hemoglobin (MCH), and Mean
Corpuscular Hemoglobin Concentration (MCHC). Together, they assist in the classification and
diagnosis of different types of anaemia and other hematological disorders.
In this practical, the blood indices were calculated using the following relationships derived from
the hematological parameters obtained during previous experiments:
MCV (fL) = (PCV × 10) / RBC count
MCH (pg) = (Hb × 10) / RBC count
MCHC (g/dL) = (Hb × 100) / PCV
The obtained values were within normal ranges:
MCV: 80–96 fL
MCH: 27–33 pg
MCHC: 32–36 g/dL
These results indicate normocytic, normochromic red blood cells, suggesting normal
erythropoietic function and hemoglobin synthesis in the subjects tested.
Variations from normal values provide diagnostic clues:
Low MCV and MCH (microcytic, hypochromic anaemia): often seen in iron deficiency or
thalassemia.
High MCV (macrocytic anaemia): occurs in vitamin B₁₂ or folate deficiency.
Low MCHC: may indicate hypochromia, while high MCHC can occur in hereditary
spherocytosis.
The practical emphasized the clinical relevance of hematological calculations in interpreting
blood disorders. While automated hematology analyzers now compute these indices instantly,
understanding their derivation enhances the student’s appreciation of the relationships between
hematocrit, hemoglobin concentration, and RBC count.
This experiment deepened our understanding of quantitative hematology, illustrating how
numerical data reflect the morphological and functional characteristics of erythrocytes, which are
crucial for diagnosing and managing anaemias.