UNIT - IV

CLINICAL LAB TECHNOLOGY

INTRODUCTION

In the era of modern technology, health care delivery system involves so many different
personnel and specialties that the caregiver must have an understanding and working knowledge
of other professional endeavors, including the role of diagnostic evaluation. Basically, laboratory
and diagnostic tests are tools by and of themselves, they are not therapeutic. In conjunction with
a pertinent history and physical examination, these tests can confirm a diagnosis or provide
valuable information about a patient status and response to therapy. In addition to these,
laboratory findings are essential for epidemiological surveillance and research purposes.

If the entire network of a laboratory service is to be effectively utilized and contribute to health
care and disease prevention, every member of its work force need to:

 Understand the role of the laboratory and its contribution to the nation’s health service;
 Appreciate the need to involve all members in the provision of health service;
 Follow professional ethics and code of conduct;
 Experience job satisfaction and have professional loyalty.

Medical laboratory science is a complex field embracing a number of different disciplines such
as Microbiology, Hematology, Clinical Chemistry, Urinalysis, Immunology, Serology,
Histopathology, Immunohematology and Molecular biology and others.

Introduction to Medical Laboratory Technology is a basic course that equips the student with the
most essential knowledge and skill pertaining to medical laboratories such as:

 Importance of laboratory services;

 Role of medical laboratory technologist;
 Use of laboratory wares, instruments and sterilization techniques;
 Prevention and control of laboratory accidents and;
 Institution of quality control system.

Moreover, this subject is extremely important for the student as it paves the ways to easily
understand various professional courses such as Hematology, Bacteriology, Urinalysis,
Parasitology, and others.

Hence, great emphasis should be given to this subject matter so as to train qualified, competent
and task oriented medical laboratory technologists.

Clinical Laboratory is a place that is equipped with different instruments, equipments and
chemicals (reagents) etc., for performing experimental works, research activities and
investigative procedures.

Medical laboratory is one part of the laboratory that is equipped with various biomedical
instruments, equipments, materials and reagents (chemicals) for performing different laboratory
investigative activities by using biological specimens (whole blood, serum, plasma, urine, stool,
etc).
Collection of Blood

Collecting blood samples and other biological specimens is crucial to the understanding,
prevention, and treatment of disease. However, from the patient’s perspective, it can also be
painful, unnerving, frightening, and inconvenient.

What blood specimen collection procedure is used depends at least in part on the nature of the
condition being tested for, but most commonly involves inserting a needle into a vein, referred to
as a "venipuncture" blood draw. Usually, medical practitioners will draw venipuncture blood
samples in their clinic offices or labs, at times convenient for them (if not always for patients).

Three popular methods of blood collection or sampling are:

1. Arterial Sampling
2. Fingerstick Sampling
3. Venipuncture Sampling

1. Arterial Sampling

This form of blood collection most commonly takes place in a hospital environment. It is
used to identify metabolic, respiratory, and mixed acid-base disorders where CO2 levels
require understanding or monitoring.

While generally safe, the procedure can be upsetting and painful for the patient. In
addition, several potential contradictions can affect the collection site, such as an
abnormal modified Allen test or local infection. There is also an increased risk of
bleeding complications in patients with coagulopathy.

2. Fingerstick Sampling

Fingerstick or finger-prick sampling involves taking a minimal amount of blood from the
patient, usually from the fingertip. Fingerstick sampling is over quickly and requires very
little preparation, which helps to reduce stress and anxiety in patients, particularly in
children and nervous adults.

Patient comfort and welfare at the point of collection is not the only reason this method
should be considered the best way to collect a blood sample. This type of sampling takes
only a few drops of blood to produce a "microsample." The long-term benefits of
microsampling to the patient include the loss of less blood and the ability to carry out
testing at home. Since fingerstick sampling is easy to do, a phlebotomist is not required
for the procedure.

Fingerstick sampling is designed to deliver a small, dried blood sample rather than a
liquid (wet) blood sample. The data derived from dried blood samples typically correlates
to the data derived from wet blood samples. Dried blood sampling can replace
conventional wet blood sampling in many scenarios. Further, since dried blood samples
 don't require cold shipping, they have the advantage of being easier and cheaper to
transport to a laboratory for analysis.

Arterial and venipuncture sampling are still widely used, and these approaches still have
their places in medicine, clinical research, and patient care. However, with advances in
technology and a greater understanding of dried blood sampling, fingerstick collection is
gaining ground.

Microsampling devices from innovators like Neoteryx, the microsampling brand of

Trajan Scientific and Medical, continue to advance blood collection and sampling. Their
solutions, including the Mitra device and the hemaPEN device, enable sample ID
tracking using a barcode system, making any mix-up or loss of samples less likely.
Technological advances in dried sampling also mean reduced contamination risks and
reduced sample rejection rates, both of which help to reduce costs. From preclinical
research to clinical trials to remote patient monitoring, the future of microsampling is
here.

3. Venipuncture sampling
Collection of urine
Collection of Stool

Collection:
 Use the large collection container provided that fits underneath the toilet seat to collect
the stool.
 Do not contaminate the stool sample with urine.
 After the stool has been collected, follow the instructions for preservation of the stool for
each of the requested tests.
 Do not remove any liquid preservative if seen in a storage container.

If collecting a stool specimen from a patient who wears diapers:

 Do not send the diaper to the laboratory.
 Remove the stool from the diaper with a disposable item like a plastic spoon or clean
unused popsicle stick.
 If the stool is more liquid than solid, then place plastic wrap inside the diaper to obtain as
much of the liquid specimen as possible.

If unable to collect enough stool for the preservative containers (explained below), then do one
of the following:

 Place the stool in a clean dry container and bring to lab as soon as possible on day of
collection. The lab may still be able to perform some if not all tests.
 Collect the stool throughout the day and combine them to increase the amount of
specimen up to the requirement.
 But do not combine specimens from different days into one specimen container.

After collection, the samples should be delivered to the laboratory as soon as possible. To
prevent leaks, make sure all lids are on tightly and that the containers are transported upright
inside a plastic bag. Make sure to bring the order slip with the specimen if the laboratory does
not already have a copy. Also, the laboratory will need insurance information and photo
identification to register the patient before testing can be done.

Methods of Collection of sputum

A sputum Gram’s stain is a laboratory test to diagnose a bacterial infection in respiratory tract.
They may order it if have symptoms of a respiratory infection that might be caused by bacteria.
It’s the most common preliminary test beyond a chest X-ray for pneumonia and other respiratory
infections, and can help promptly prescribe a treatment plan. To complete a sputum Gram’s
stain, will need to a collect a sample of sputum and send it to a laboratory for testing. Sputum is a
mixture of saliva and mucus that you cough up from your respiratory tract. It’s usually colored
and thick in consistency, indicates have an infection in lungs.

Plain saliva comes from mouth and is usually clear. The collection of sputum can be done by
four ways, which includes:
 1. Spontaneous sputum collection: patients coughs up sputum into a sterile container. The
advantage of this method is inexpensive and easy to do. The disadvantages are patient
may not be able to cough up sputum without assistance or may spit up saliva instead.
Healthcare worker has to coach and supervice the patient when collecting sputum.

2. Sputum induction: the patient inhales a saline mist which can cause a deep cough. This
method is to do and use to obtain sputum when coughing sputum is not productive. The
disadvantage of this method is specimens may be watery and confused with saliva
(should be labeled with induced specimen) and requires special equipment and may cause
bronchospasam.

3. Bronchoscopy: the bronchoscope is passed through the mouth or nose directly into the
diseased portion of the lung, and sputum or lung tissue is removed. The use of this
method is to obtain sputum when coughing sputum is not productive or other diagnoses
are being considered. The disadvantages of this method are most expensive and invasive
procedures, requires special equipment, must be done by a specialized physician in a
hospital or clinic and requires anesthesia.

4. Gastric washing: In this method the tube is inserted though the patients mouth or nose
and passed into the stomach to get a sample of the gastric secretions that contain sputum that has
been coughed into the throat and then swallowed. The advantage of this method is use to obtain
samples in children, who do not produce sputum when they cough. The disadvantage of this
method is must be done as soon as the patient wakes up in the morning, patient may be required
to stay in the hospital and can be uncomfortable for the patient.

Storage of clinical samples

A modern analytical laboratory handles several hundred samples in a single day. Sample
management plays a crucial role as without a sample management plan in place it will not be long
before the laboratory presents a chaotic look with samples lying all around. Valuable time will be
lost in trying to locate the required samples.

Sample Storage

Proper sample storage and prevention of cross contamination are the main considerations after
receipt of samples in the laboratory. Some suggested guidelines are provided here.

 Sample labels should be legible and include sample details. Such details should include
sample name, batch number (if applicable), date of preparation or collection and
recommended storage conditions.
 Ensure that sample container or pouch is properly sealed and there is no leakage of material
 Samples need not be stored in an alphabetical order. It may be easy to locate samples stored
alphabetically but there are other better plans for sample storage such as storage on basis of
priority in separate bins – most urgent, urgent or normal priority. This plan may involve
movement of samples between different bins when priorities change.
  Make a manual/soft record on sample details such as sample name, code(if applicable) date
and time of receipt , sample source, analysis requirements, promised result delivery date,
actual date of delivery and finally sample status ,ie, pass or fail
 Light and temperature sensitive samples should be stored under the prescribed conditions.

Sample Disposal

Technical details for sample disposal procedures are not part of this write-up and only general
guidelines are provided. It is a good to keep in mind that samples after analysis should not be
disposed off in washbasins or dustbins.

 Maintain record of disposal of samples similar to the ones maintained for samples before
analysis
 Retain sufficient quantity of sample under recommended storage conditions for the purpose
of retesting for verification of results in case of disputes or confirmation of results
 Engage special agencies in case of hazardous sample disposal
 Disposal of samples should not pose any hazard to other laboratory workers or visitors to
the laboratory so it is absolutely essential to adopt recommended practices for storage and
disposal of samples.

The general guidelines offered provide useful tips but can devise sample storage and disposal plans
based on nature of samples handled and daily workloads.

Techniques of preservation of samples

Proper preservation practices must be followed and can be found for each analyte in a clinical
sample. Following collection and during transportation, samples should be kept at 60C or on ice.
Samples requiring preservation should be preserved as soon as possible after collection to
maintain the integrity of the sample.

Methods of preservation are intended to retard biological action, retard hydrolysis of chemical
compounds and complexes, and reduce volatility of constituents. Preservation methods are
limited to pH control, chemical addition, amber or opaque bottles, filtration, refrigeration, and
freezing.

To minimize the potential for volatilization or biodegradation between sampling and analysis,
keep the samples as cool as possible without freezing. Preferably, pack samples in crushed or
cubed ice, or a commercial ice substitute before shipment. Avoid using dry ice because it may
freeze the samples and cause the containers to break. Dry ice may also affect the pH of the
samples. Keep composite samples cool with ice or a refrigeration system set at 6 0C during
collection. Analyze the samples as quickly as possible upon arrival at the laboratory. If
immediate analysis is not possible, storage at 20-40C is recommended for most samples.

Use chemical preservation only when it is shown not to interfere with the method of analysis.
When chemicals are used, add them to the sample bottles so that all the sample portions are
evenly preserved as soon as collected. No single preservation method is entirely satisfactory;
choose the preservation with regard to the analyses being made. Because a preservation method
for one analysis may interfere with the preservation for another, samples for multiple
determination may need to be split and preserved separately. All methods of preservation may be
inadequate when applied to suspended matter. DO NOT use formaldehyde as a preservative
because it affects numerous analyses.

Separation of blood plasma and serum

Serum Preparation from Blood

Collect whole blood in a microcentrifuge tube. After collection of the whole blood, allow the
blood to clot by leaving it undisturbed at room temperature. This usually takes 15-30 minutes.
Remove the clot by centrifuging at 1,000-2,000 x g for 10 minutes in a refrigerated centrifuge.
The resulting supernatant is designated serum. Following centrifugation, it is important to
immediately transfer the liquid component (serum) into a clean microcentrifuge tube using a
pipette.

The samples should be maintained at 2-8°C while handling. If the serum is not analyzed
immediately, the serum should be stored and transported at –20°C or lower. It is important to
avoid multiple freeze-thaw cycles because this is detrimental to many serum components.
Samples that are hemolyzed, icteric, or lipemic can invalidate certain tests.

Plasma Preparation from Blood

Collect whole blood into commercially available anticoagulant-treated tubes e.g., EDTA-treated.
Cells are removed from plasma by centrifugation for 10 minutes at 1,000-2,000 x g using a
refrigerated centrifuge. Centrifugation for 15 minutes at 2,000 x g depletes platelets in the
plasma sample.

The resulting supernatant is designated plasma. Following centrifugation, it is important to

immediately transfer the liquid component (plasma) into a clean microcentrifuge tube using a
pipette. The samples should be maintained at 2-8°C while handling. If the plasma is not analyzed
immediately, the plasma should be stored and transported at –20°C or lower. It is important to
avoid multiple freeze-thaw cycles. Samples which are hemolyzed, icteric, or lipemic can
invalidate certain tests.

Blood smear Preparation

Blood collection for thick and thin blood smears

Capillary blood obtained by fingerstick:

1. Label pre-cleaned slides (preferably frosted-end) with patient’s name (or other identifier),
date and time of collection.
2. Wear gloves.
 3. Clean slides with 70 to 90% alcohol and allow to dry. Do not touch the surface of the
slide where the blood smear will be made.
4. Select the finger to puncture, usually the middle or ring finger. In infants, puncture the
heel.
5. Clean the area to be punctured with 70% alcohol; allow to dry.
6. Puncture the ball of the finger, or in infants puncture the heel.
7. Wipe away the first drop of blood with clean gauze.
8. Touch the next drop of blood with a a clean slide. Repeat with several slides (at least two
thick and two thin smears should be made). If blood does not well up, gently squeeze the
finger.

For venous blood obtained by venipuncture:

1. Label collection tubes and pre-cleaned slides (preferably frosted-ewith the patient’s name
(or other identifier), date and time of collection.
2. Clean the site for blood collection well using 70% alcohol; allow to dry.
3. Collect the venous blood in a vacuum tube containing anticoagulant (preferably EDTA);
alternatively, collect the blood in a syringe and transfer it to a tube with anticoagulant;
mix well.
4. Prepare at least two thick smears and two thin smears as soon as possible after collection.

Making thick and thin blood smears

1. Whenever possible, use separate slides for thick and thin smears.
2. Thin film (a): Bring a clean spreader slide, held at a 45° angle, toward the drop of blood
on the specimen slide.
3. Thin film (b): Wait until the blood spreads along the entire width of the spreader slide.
4. Thin film (c): While holding the spreader slide at the same angle, me angle, push it
forward rapidly and smoothly.
5. Thick film: Using the corner of a clean slide, spread the drop of blood in a circle the size
of a dime (diameter 1-2 cm). Do not make the smear too thick or it will fall off the slide.
6. Wait until the thin and thick films are completely dry before staining. Fix the thin film
with methanol (100% or absolute) and let it dry completely before staining. The thick
film should not be fixed.
7. If both thin and thick films need to be made on the same slide, fix only the thin film with
methanol. The thick film should not be fixed.

Erythrocyte Sedimentation Rate (ESR) and Clinical Significance:

The Erythrocyte sedimentation rate (ESR) is a common hematological test for nonspecific
detection of inflammation that may be caused by infection, some cancers and certain
autoimmune diseases, it can be defined as the rate at which Red Blood Cells (RBCs) sediment in
a period of one hour.
Principle

When anticoagulated blood is allowed to stand in a narrow vertical glass tube, undisturbed for a
period of time, the RBCs – under the influence of gravity – settle out from the plasma. The rate
at which they settle is the measured as the number of millimeters of clear plasma present at the
top of the column after one hour (mm/hr). This mechanism involves three stages.

 Stage of aggregation: It is the initial stage in which pilling up of RBCs takes place. The
phenomenon is known as Rouleaux formation. It occurs in the first 10-15 minutes.
 Stage sedimentation: It is the stage of actual falling of RBCs in which sedimentation
occurs at constant rate. This occurs in 30-40 minutes out of 1 hour, depending upon the
length of the tube used.
 Stage of packing: This is the final stage and is also known as stationary phase. In this,
there is a slower rate of falling during which packing of sedimented RBCs in column
occurs due to overcrowding. It occurs in final 10minutes in 1hour.

Methods of ESR determination

There are two main methods to determine ESR:

1. Wintrobe’s method
2. Westergren’s method

Each method produces slightly different result. Mosely and Bull (1991) concluded that
Wintrobe’s method is more senisitive when the ESR is low, whereas, when the ESR is high, the
Westergren’s method is preferably an inidication of patients clinical state.

1. Wintrobe’s method:
This method uses wintrobe’s tube, a narrow glass tube closed at the lower end only. The
Wintrobe’s tube has a length of 11 cm and internal diameter of 2.5 mm. It contains 0.7-
1ml of blood. The lower 10 cm are in cm and mm. The marking is 0 at the top and 10 at
the bottom for ESR. This tube can also be used for PCV. The marking is 10 at the top and
0 at the bottom for PCV.

Requirements:

 Anticoagulated blood (EDTA, double oxalate)

 Pasteur pipette
 Timer
 Wintrobe’s tube
 Wintrobe’s stand

Procedure

1. Mix the anticoagulated blood thoroughly.

2. By using Pasteur pipette, fill the Wintrobe’s tube upto ‘0’ mark. There should be no
bubbles in the blood.
 3. Place the tube vertically in ESR stand and leave undisturbed for 1 hour.
4. At the end of 1 hour, read the result.

Normal values:

For males : 0 -9 mm/hr.

For females: 0-20mm/hr.

WESTERGREN’S METHOD

It is the better method than Wintrobe’s method. The reading obtain is magnified as the column is
lengthier. The Westergren tube is open at the both ends. It is 30 cm in length and 2.5 mm in
diameter. The lower 20 cm are marked with 0 at the top and 200 at the bottom. It contains anbout
2 ml of blood.

Requirements

 Anticoagulated blood (0.4ml of 3.13% trisodium citrate solution + 1.6 ml of blood).

 Westergren tube
 Westergren stand
 Rubber bulb (sucker)

Procedure

1. Mix the anticoagulated blood thoroughly

2. Draw the blood into the tube upto 0 mark with help of rubber bulb.
3. Wipe out blood from bottom of the tube with cotton.
4. Set the tube upright in stand. Make sure the pipettes fits snugly to eliminate possible
leakage and that the pipette is in vertical position.
5. Leave the tube undisturbed for 1 hour.
6. At the end of 1 hour, read the result.

Normal values

For males : 0 -10 mm/hr

For females: 0-15 mm/hr.

Clinical significance:

The erythrocyte sedimentation rate (ESR) is non-specific test. It is raised in wide range fo
infectious inflammatory, degenerative, and malignant conditions associated with changes in
plasma proteins, particularly increases in fibrinogen, immunoglobulins, and C-reactive protein.
The ESR is also affected by many other factors including anaemia, pregnancy,
heamoglobinopathies, haemoconcentration and treatment with anti-inflammatory drugs.

Causes of significantly raised ESR

 All types of anemias except sickle cell anemia

 Acute and chronic inflammatory conditions and infections including: HIV disease,
Tuberculosis, Acute viral hepatitis, Arthritis, Bacterial endocarditis, Pelvic inflammatory
disease, Ruptured ectopic pregnancy,
 Systemic lupus erythematosus,
 African trypanosomiasis,
  Visceral leishmaniasis,
 Myelomatosis, Lymphoma, Some tumors,
 Drugs including oral contraceptives.

Causes of reduced ESR

 Polycythaemia
 Poikilocytosis
 Newborn infants
 Dehydration
 Dengue haemorrhagic fever
 And other conditions associated with haemoconcentration.

Method of Determination of PCV (or ) Packed Cell Volume

When anti-coagulated blood is centrifuged at a standard speed, erythrocytes, which are heavier
than white cells and plasma, will settle down at bottom. This red cells volume is known as
Haematocrit or Packed Cell Volume (PCV). Haematocrit or PCV is the volume of red cells
expressed as a percentage of whole blood.

Methods:

There are two methods, used for the determination of haematocrit:

1. Macrohaematocrit
2. Microhaematocrit

1.Macrohaematocrit:
A large volume of blood is required in this method. Approximately 2 to 4 ml is required.

Principle:

Anticoagulated blood is taken in a Wintrobe tube. Fill upto the uppermost mark and then rotate
for desired length of time. The packed cell volume (PCV) of red cells is directly read from the
graduated tube as %.

Requirement:

1. Blood specimen:EDTA or double oxalated anti-coagulated blood is used in this method.

Determine P.C.V. within six hr. of blood collection.
2. Wintrobe Tube:It is 110 mm in length and 2.5 mm in diameter. The lower 100 mm are
graduated or marked, from 100 at top and 0 (zero) at bottom for PCV.
3. Long necked pasture pipette or a special type of syringes is used for filling Wintrobe
tube.
4. Centrifuge machine with known speed.
Procedure:

Mix 0.4 ml of EDTA with 2 ml blood. Fill the Wintrobe tube upto upper most mark with the help
of pasture pipette or syringe. Fill the another Wintrobe tube to balance first one. If the blood
sample is not available, fill the tube with water.

Place the Wintrobe tube in opposite side in centrifuge. Turn the centrifuge to slow speed, then
slowly increase the speed to 3,000 rpm. Centrifuge for 30 min. at 3,000 rpm. After 30 min.
switch off the centrifuge and allow it to stop by itself. Take out the Wintrobe tube and read PCV
directly with the help of graduation mark given on the tube.

Normal Value:
 In male – 42 to 50%
 In female – 36 to 38%.

Microhaematocrit:

This method requires small amount of blood, 2 to 3 drops only. The blood can be obtained by
finger puncture.

Principle:

Anti-coagulated blood is centrifuged in a sealed capillary tube, and then PCV is determined by a
special haematocrit reader.

Requirement

1. Blood Specimen: Blood from finger puncture may be used or EDTA or double oxalate
venous blood can also be used.
2. Capillary Tube: Use plain capillary tube for anti-coagulated venous blood and use
heparinised capillary tube (Coated with heparin internally) for blood obtained from finger
puncture. The capillary tube is approximately 75 mm in length.
3. Microhaematocrit Centrifuge: This is a special type of centrifuge. It has speed about
15,000 r.p.m. The top of centrifuge is flat with grooves. The centrifuge also has timer,
which is usually set for 5 min.
4. Haematocrit Reader: There are several types of readers used for reading hct. The
simplest method is use of card reader, which can be made by hand.
5. Clay: This is used to seal the end of capillary tube.

Procedure:

Draw the blood sample into appropriate capillary tube with capillary action. Use plain tube for
anti-coagulated blood and heparinised tube for plain blood. In case of finger puncture, the blood
should flow freely with little pressure. Now wipe off the first drop and then collect the blood
specimen.
Fill the tube about 3/4th length with blood. Seal the another end of the tube with clay or wax or
ultimately by heating. The sealing should be about 2 mm deep. Place two hct tubes in the groove
of centrifuge exactly opposite to each other.

It is not necessary that the capillary tube have exact amount of blood level. In case, if there is no
filled capillary tube to balance we can use an empty capillary tube. Centrifuge at 13,000 ± 2000
rpm.

Remove capillary tube from centrifuge. It will show three layers. Top layer is of plasma or
serum; the middle layer is thin creamy white in colour and is known as Buffy coat. It is a layer of
WBC; the last layer is the column of RBC. Use the hct reader for finding the value of hct.

Card Reader:

The reader is used as, hold the tube against Scale so that the bottom of red cell is matched with 0
(zero) line of the card. Move the tube across the card until the uppermost line of plasma is
matched to 100% line of card.

Check to make sure that bottom of red cell column is still in the line of zero and the tube should
be straight and vertical. The line that passes through the top of the column of RBC gives the hct
Value.

Importance:

 Decrease in PCV is suitable measure for anaemia. The fall in hct may be seen in
decreased oxygen supply to cells, heart disease or malignant condition, hct also rise in
case of dehydration.
Blood Indices

Blood or Red blood cell (RBC) indices measure the size, shape, and quality of red blood cells.
Red blood cells, also known as erythrocytes, carry oxygen from lungs to every cell in our body.
Cells need oxygen to grow, reproduce, and stay healthy.

There are four types of red blood cell indices:

• Mean corpuscular volume (MCV), which measures the average size of red blood cells.
Mean corpuscular volume (MCV) is the average volume of a red blood cell and is
calculated by dividing the hematocrit (Hct) by the concentration of red blood cell count.

• Normal range: 80–100 fL (femtoliter).

• Mean corpuscular hemoglobin (MCH), which measures the average amount of

hemoglobin in a single red blood cell. Hemoglobin is a protein in red blood cells that
carries oxygen. Mean corpuscular hemoglobin (MCH) is the average amount
of hemoglobin (Hb) per red blood cell and is calculated by dividing the hemoglobin by
the red blood cell count.

• Normal range: 27-31 pg/cell

• Mean corpuscular hemoglobin concentration (MCHC), which also measures

hemoglobin in red blood cells. In addition, it includes a calculation of the size and
volume of red blood cells. Mean corpuscular hemoglobin concentration (MCHC) is the
average concentration of hemoglobin per unit volume of red blood cells and is calculated
by dividing the hemoglobin by the hematocrit.

• Normal range: 32-36 g/dL

• Red cell distribution width (RDW), which measures differences in the volume and size
of red blood cells. If one or more of these indices are not normal, it may mean have some
type of anemia, a condition in which body does not make enough healthy red blood cells.
Uses of blood indices

Red blood cell (RBC) indices are part of a complete blood count, a group of tests that measures
various parts and features of blood. The results of RBC indices are used to diagnose different
types of anemia. There are several types of anemia, and each type has a different effect on the
size, shape, and/or quality of red blood cells.

Red blood cell indices testing needs

If have any symptoms of anemia, which include:

• Weakness
• Fatigue
• Pale skin
• Rapid heartbeat
• Cold hands and feet
• Shortness of breath

Red blood cell indices test

A health care professional take a blood sample from a vein of arm, using a small needle. After
the needle is inserted, a small amount of blood will be collected into a test tube or vial. This test
usually takes less than five minutes.

Interpretation of results from test

Abnormal results may include one or more of the following:

Mean corpuscular volume (MCV)

If red blood cells are smaller than normal, it may mean have:
• Iron deficiency anemia, the most common form of anemia. It happens when don't have
enough iron in body.
• Thalassemia, an inherited disease that can cause severe anemia.

If red blood cells are larger than normal, it may have:

• Anemia caused by a vitamin B deficiency
• Liver disease

Mean corpuscular hemoglobin (MCH)

If the amount of hemoglobin is lower than normal, it may mean have:

• Iron deficiency anemia

If the amount of hemoglobin is higher than normal, it may mean have:

• High level of cholesterol in the blood

• Anemia caused by a vitamin B deficiency
Mean corpuscular hemoglobin concentration (MCHC)

If the average amount of hemoglobin is lower than normal, it may mean have:
• Iron deficiency anemia
• Thalassemia

If the average amount of hemoglobin is higher than normal, it may have:

• Hemolytic anemia, a type of anemia that happens when red blood cells are broken up in
the blood stream
• Hereditary spherocytosis, a rare genetic disorder that causes anemia and gallstones

Red cell distribution width (RDW)

If results were normal, it means red blood cells are normal in size and are all about the same size.
If results were not normal, it means there are differences in the size of red blood cells. This
measurement is not enough to make a diagnosis, so RDW results are usually combined with the
results of other indices and other blood tests. The combination of results can help confirm a
diagnosis.

Platelet count

RBC pipette is used to do the platelet count

Thrombocytopenia is a condition in which have a low blood platelet count. Platelets
(thrombocytes) are colorless blood cells that help blood clot. Platelets stop bleeding by clumping
and forming plugs in blood vessel injuries.

Thrombocytopenia might occur as a result of a bone marrow disorder such as leukemia or an

immune system problem. Or it can be a side effect of taking certain medications. It affects both
children and adults.

Thrombocytopenia can be mild and cause few signs or symptoms. In rare cases, the number of
platelets can be so low.

Symptoms

Thrombocytopenia signs and symptoms may include:

• Easy or excessive bruising (purpura)
• Superficial bleeding into the skin that appears as a rash of pinpoint-sized reddish-purple
spots (petechiae), usually on the lower legs
• Prolonged bleeding from cuts
• Bleeding from gums or nose
• Blood in urine or stools
• Unusually heavy menstrual flows
• Fatigue
• Enlarged spleen and Treatment options are available.

Bleeding Time & Clotting Time

The Bleeding and Clotting time test refers to a test that is performed on a sample of blood to
measure the time taken for it to clot or coagulate. This test is also known as the BT CT test.

In a bleeding time test, it is assessed the rapidness with which the blood can clot and it can stop
bleeding is. In this test, a small puncture is made in the skin of the person. By performing this
test, it can be easily determined the way in which the platelets work together to form clots.

In Clotting time test is the time taken for blood to clot in a person. Clotting factors determine the
clotting time in a person. Thus blood clotting factors play an important role in bleeding and
clotting.

Platelets are found in the blood. Their function is to accumulate near the site of injury or
puncture to seal the wound and reduce or stop the amount of blood that is flowing away from the
body.

Significance of BT & CT

If the patient is experiencing an issue with blood clotting wherein the blood is not able to stop
flowing after an injury such as a cut or a puncture, then it is suggested to take the bleeding time
test to determine if the person has problems with blood clotting.
Blood clotting disorders symptoms are the delay in blood clotting and longer bleeding time. The
bleeding time test is the most common test to check if the blood clotting has issues. The
blood, not clotting disease occurs in very few people.

There are also a few other tests available to evaluate if a person suffers from bleeding problems.
If there is prolonged bleeding in a person, it indicates that the person has an acquired defect of
platelet function. The test is also carried out to determine epistaxis. Blood clotting mechanism in
few people may not function properly, and hence clotting could be difficult.

Interpretation of BT & CT

The normal bleeding time is between 2-7 minutes. The normal clotting time in a person is
between 8-15 minutes. By understanding the time taken for blood to clot, it can be determined if
the person has haemophilia or von Willibrand’s disease.

Bleeding time normal range can still be considered between a 1 - 8 minutes.

If the bleeding time is outside the range, it could imply an underlying platelet defect, and there
should be more tests done to confirm it. An abnormal bleeding time indicates that the person
could have acquired platelet function defect. An acquired platelet function defect develops after
birth.

In this kind of a defect, the platelets may not be working properly, or the body might be
producing too many platelets or fewer platelets. The abnormal results could mean:

• That the person has a defect in the blood vessel wherein the blood vessels are unable to
transport blood properly throughout the body
• That the person has a genetic platelet function disorder by birth. This genetic disorder
could affect the function of the platelets. For example haemophilia.
• The person could be suffering from thrombocythemia wherein the person bone marrow
starts creating too many platelets in the body.
• The person could be suffering from thrombocytopenia wherein the person bone marrow
starts creating too little platelets in the body.
• The person may be suffering from Von Willebrand’s disease. This is an acquired
hereditary disease that affects the process of blood coagulation in a person.

Bleeding Time & Clotting Time Test procedure

The test is performed by a nurse or a doctor. The nurse/doctor cleans the site of puncture with an
antiseptic to ensure that there is a minimal infection from the procedure. A pressure cuff is
placed around the arm, and it is inflated.

The pressure cuff is placed on the upper arm. Then, two cuts that are small in size are made on
the lower arm. These cuts cause a little bleeding. These are extremely shallow cuts. Then the cuff
is removed from the arm.
The bleeding time and clot time is checked using a timer. Every 30 seconds the blood from the
cuts is blotted with blotting paper till the bleeding stops. Once the procedure is completed the
cuts are bandaged.

Results

The bleeding time test may be affected by medications that are used by the person. There are
several medications that may act as anticoagulants and affect the time taken for bleeding and
clotting. Hence if the person is on any medication or supplement or herbs, this should be
immediately informed to the doctor.

Also, a diet full of green and leafy vegetables may affect the clotting time. If the person has very
low platelet count, then the BT test should not be done on the person.

Also, people on anticoagulants or who have lymph nodes dissected should not undergo the BT
test. The scars that are made for the BT test remain visible for a long time in people who tend to
have keloids.

Results of Bleeding Time And Clotting Time

Reference Range Interpretation

2-7 minutes (Bleeding Time) Normal

8-15 minutes (Clotting Time) Normal

Examination of urine

Sample collection or urine collection is already studied in the methods of collection of

clinical samples (URINE SPECIMEN).
Microscopic urine examination
Pregnancy Tests

Pregnancy test is mainly used to check the special hormone —human chorionic gonadotropin
(HCG) — that only develops in a person’s body during pregnancy. These tests can use either pee
or blood to check for HCG. At-home pregnancy tests that use pee are the most common type.
When used correctly, home pregnancy tests are 99% accurate.

A pregnancy test is used to determine whether the individual is pregnant or not. If pregnancy test
is positive, it means pregnant. If the test is negative, it means aren’t pregnant. Pregnancy tests
work by detecting human chorionic gonadotropin (HCG), a hormone body makes when were
pregnant.

From the very beginning of pregnancy, body starts to go through changes to support the cells that
will develop into baby. One thing that happens very quickly is the production of HCG. If
pregnant, body starts to produce more HCG. HCG levels start to build up once the fertilized egg
implants in uterus — about six to 10 days after conception.

There are two main types of pregnancy tests — urine tests and blood tests.

Urine test at home with a home pregnancy test. This type of test is available over the counter and
in a variety of price ranges.
Blood tests to check for pregnancy happen in healthcare provider’s office and involve giving a
sample of blood. The other way to confirm a pregnancy is by using an ultrasound. Healthcare
provider performs an ultrasound in their office.

Pregnancy tests mainly look for an elevated amount of HCG. Levels of HCG rise quickly –
doubling every few days in the first weeks of pregnancy. The placenta produces HCG. Only
pregnant people have a placenta, which develops shortly after a fertilized egg attaches to uterine

Pregnancy tests work by reacting to the amount of HCG in either pee or blood. In a urine test, a
piece of reactive paper detects the HCG. This test might show a plus sign, double vertical lines
or even the word “pregnant.” Different tests will show a positive result in unique ways. Read the
directions that come with the test to know a positive result will look like. For example, most tests
have a control window that shows up first. Seeing a symbol in this window will explain that the
test is working. The different brands of tests will take different amounts of time to show a result.
There are three ways to take an at-home pregnancy test:

• Pee in a clean cup. Then, place one to several drops of pee on a chemical strip.
• Place the pregnancy test strip in urine stream while pee.
• Pee in a clean cup and then dip the test strip in the pee while it’s still in the cup.

For many of these tests, HCG can be detected in your urine about 10 days after conception.
However, taking it after you miss your period reduces the chance of getting a false-negative
result. A missed period typically happens around 14 days after conception.

Blood test

Another type of pregnancy test is a blood test. Blood tests are rarely done because they’re
expensive and tend to have the same result as a urine test. This type of pregnancy test is done
using a small sample of blood from a vein in arm. This blood test not only detects whether the
pregnancy hormone is in our body, but can also determine how much of the hormone is present.
This is helpful for when provider needs to know the exact amount of HCG in blood, not just if
there’s HCG in blood.

A blood test for pregnancy might be done in special circumstances, such as for people who are
having fertility treatments or when the healthcare provider thinks there might be a problem.
These blood tests are slightly more sensitive than urine tests because they can detect very small
levels of HCG. That means they can provide a more accurate answer very early on in pregnancy
— within seven to 10 days after conception. For this test, blood sample is taken at provider’s of
hospital, then sent to a lab for analysis. Results might take anywhere from a few hours to two
days.
.
A blood test confirms pregnancy first because it can detect a smaller amount of HCG as
compared to a test that uses pee.
Examination of stool or fecal matter