HEMATOLOGY 1 BSMLS2

MIDTERMS | LABORATORY

Prof. John Daryll R. Nacua, RMT

refers to the process of obtaining a sample of blood from an

individual for diagnostic, screening, or therapeutic purposes.
It is typically performed by a healthcare professional, such as
a nurse or phlebotomist, using specialized techniques and
equipment.

The collected blood sample can be used for various medical

tests, including blood cell counts, glucose levels, cholesterol
levels, infectious disease screening, genetic testing, and many
other diagnostic procedures. Blood collection is a crucial
component of healthcare as it provides valuable information
for diagnosing and monitoring a wide range of medical VENIPUNCTURE
conditions. ● Ideal procedure is to have the patient lie down
or if not possible, the patient should sit in a
sturdy, comfortable chair and not on high stools.
THE TYPES OF BLOOD COLLECTION
● There should be nothing inside the patient's
Venipuncture mouth during the procedure.

Capillary Puncture ● Ideal site for puncture: Antecubital Fossa

○ Median Cubital Vein
Arterial Puncture ○ Cephalic / Median Cephalic Vein
○ -Basilic Vein

A MUST IN PHLEBOTOMY:

1. Correct patient identification.

2. Correct specimen identification. (Proper

labeling: Patient's Full Name, Hospital
Identification Number, Location, Time and Date
and the initials of the phlebotomist)
3. Consistent with universal precautions (Gloves,
Gowns must be worn at all times and hands
must be washed in between patients)

4. Aseptic Technique (Cleaning of puncture site

“H - PATTERN”
with 70% alcohol)
1. Median Cubital Vein
2. Cephalic Vein
5. Sharp objects must be thrown to appropriate 3. Basilic
containers and must not be unsheathed or bent.
“M - PATTERN”
6. Gauze and cotton must be waste in biohazard 1. Median Vein
containers or trash bins. 2. Accessory Cephalic Vein
3. Basilic
Transcribed by: Don Filomeno Marfori B. Agus

● Assemble the materials.

● Identify and position the patient.

● Less than 30°: Angle between skin and needle
● Less than 1 minute: Tourniquet Application ● Apply a tourniquet 2-4 inches away from the
● Hemoconcentration, Hemolysis, Shortened site of the puncture.
Coagulation Time (PT/APTT) are the possible
effects of prolonged tourniquet application. ● Select the vein for puncture. Release
● Application of tourniquet must be 3-4 inches tourniquet.
above the puncture site.
● a 21 gauge (1 inch long) needle is the most ● Apply antiseptics (70% alcohol) in a circular
common needle size for adult. manner starting from inside then out. Let it
● Phlebotomist must never puncture the patient dry.
twice. ● Reapply tourniquet. Insert needle with 15-30
● Patient should not pump the fist. degrees angle

● Withdraw blood by pulling the plunger, not

too fast, not too slow

● Release tourniquet

● Withdrawing the needle. Put cotton on the

puncture site then apply pressure.

● Transfer blood sample to anticoagulated tube.

Remove needle and tube cap and let blood
flow on the sides to avoid hemolysis.

Venipuncture: Closed Method

● Prolonged tourniquet application Moisture or

contamination of blood collecting tubes.

● Needle with small bores.

● Excessive agitation.

● Frothing of the blood sample.

● Assemble vacutainer equipment.

● Identify the patient and position the patient's

Starting from the left: (1) hemolyzed sample, (2) Lipemic forearm.
sample, (3) Normal sample, (4) Icteric sample.
● Apply a tourniquet, 2-4 inches above the
Venipuncture: Open Method puncture site.

● Select the best vein. Then release tourniquet

.
● Cleanse the area with proper aseptic
technique. Then reapply the tourniquet.

● Using a free hand, stabilize the vein.

● Advance needle through the skin (15-30

degrees angle). Stabilize the vacutainer with
- the thumb top and three fingers underneath.
Transcribed by: Don Filomeno Marfori B. Agus

● Keeping the holder steady, gently push the

tube forward.

● Release tourniquet. Remove vacutainer tube

and then withdraw the needle.

● Apply pressure on the site with a dry cotton

ball.

● Perform under supervision (students)

● Be sure that the patient is in the proper position

and be prepared that the patient will faint.

● Do not allow the tourniquet to remain in

position for more than one minute.

● Before inserting the needle, push the plunger all

the way to expel air.

● Always remove the tourniquet before removing

the needle.

● Do not reuse needles or syringes

● Proper waste disposal.

Capillary / SKin Puncture

Capillary puncture is done if the patient is:

● Infants less than 1 year old.

● Severely burned patients.

● Patients whose veins are reserved for

therapeutic purposes.

● Extremely obese patients.

● Adult with poor veins.

Transcribed by: Don Filomeno Marfori B. Agus
● Mastectomy Patients
● Avoid applying pressure, squeezing, "milking"
○ Draw blood from the opposite of
.
mastectomy side.
● Order of Draw for
○
○ Tube for blood gas analysis
● Obesity
○ Slides
○ BP Cuff will help
○ EDTA Microcollection Tube
■ Must not be more than 40
○ Other Microcollection Tube with
mmHg and 1 minute
anticoagulants
■
○ Serum Microcollection Tube
● iatrogenic Anemia
● Discard first drop of blood ● Failure to Draw Blood
○ Discard excess tissue fluid
○ Discard dead epidermal cells ● Petechiae
○ Facilitate free flow of blood
● Nerve Damage
● Depth of Skin Puncture
● Hemolysis
○ Adult: 2.0-2.5 mm
○ Infants: <2.0 mm ● Burned, Damaged, Scarred ad Occluded Veins

● -Seizures and tremors

Materials for Capillary Puncture ● Vomiting and Choking

● Blood lancet ● Allergies

● Cotton Balls
● 70% Alcohol ● Dialysis Patients
● Capillary Tubes
● Gauze ● Edema
● Sealing Clay

● Hematoma
● Burned, damaged, occluded veins
● Intravenous catheter (IV line)
● Edema
● Post Mastectomy Side
● Skin with Tattoo

Complications during Venipuncture

● Ecchymosis (Bruise)
○ Most common
○ Leakage of small amount of blood

● Hematoma
○ Leakage of large amount of blood.

● Fainting (Syncope)
○ Short lapse in consciousness

● Hemoconcentration
○ Prolonged tourniquet application
○ Wait for 2 minutes before reapplying
the tourniquet

● Intravenous (IV) Therapy

○ Draw in the opposite side of the IV
○ Stop IV for 2 minutes
○ Discard first 5-ml of blood
Transcribed by: Don Filomeno Marfori B. Agus
● Reversible (Treated by administration of
Ascorbic Acid and Methylene Blue

Hemoglobin (HgB)
B. Sulfhemoglobin
● Also known as Respiratory Pigment.
● Addition of Hydrogen sulfide to the hemoglobin
● One of the tests used to diagnose and follow (greenish pigment)
the treatments of anemia.
● Irreversible
● Main component of the RBC (95%)
● caused by sulfonamides, phenacetin, nitrites,
● Carry oxygen to and carbon from tissues. phenylhydrazine
● Heme: has iron (in ferrous state: binds 02)
C. Carboxyhemoglobin
● Globin: portion (determines the type of
hemoglobin)Each Hgb has 4 subunits: 4 heme & ● combination of heme and carbon monoxide
4 globin
● CO has an affinity to hemoglobin of 240x than
● 1 heme is capable of carrying 1 mole of O2: 1 that of oxygen.
Hgb can carry moles of O2.
● reversible (Treated by Oxygen Saturation or
Hyperbaric oxygen therapy)

Types of Hemoglobin (Normal)

Hemoglobin A (2 alpha chains/2 beta chains) > 95%

Hemoglobin A2 (2 alpha chains/2 delta chains) < 3.5% ● Reference Method: Cyanmethemoglobin
(Hemiglobincyanide)

● Uses Drabkin's Reagent

Hemoglobin F (2 alpha chains and 2 gamma chains) 1-2%
○ Contains Potassium Cyanide, Potassium
Ferricyanide, potassium dihydrogen
● Alpha chain is coded by Chromosome 16 phosphate or sodium bicarbonate, and
● Gamma, delta and beta by Chromosome 11
a non ionic compound (improves cell
lysis)
Types of Hemoglobin (Abnormal)

● Hemoglobin S Potassium ferricyanide: hemoglobin → methemoglobin

● Hemoglobin C
● Hemoglobin E Potassium cyanide: methemoglobin → cyanmethemoglobin

● Hemoglobin D
● Hemoglobin G
● Hemoglobin Lepore ● Used EDTA Anticoagulated blood

● All hemoglobin types and variants are measured

except sulfhemoglobin.

● Read 540 nm

➢ Hemoglobin whose structures have been ● 5 ml or 5000 ul of reagent, 20 ul or 0.02 ml of

modified due to drugs or environmental whole blood.
chemicals. They are dysfunctional hemoglobin
that are unable to transport oxygen.
REFERENCE RANGE (RODAK)
A. Methemoglobin
● Male 14 - 18 g/dL
● Can cause Increased O2 affinity thus decreasing
the efficiency of Oxygen to be delivered to the ● Female 12 - 15 g/dL
tissues. ● Newborn 16.5 - 21.5 g/dL

● Caused by: oxidants (Nitrite, Primaquine,

dapsone, Benzocaine) or Methemoglobin
Reductase deficiency
Transcribed by: Don Filomeno Marfori B. Agus

Technical
● Pipetting errors
● Dirty and scratch cuvettes
● Deteriorated reagent

Biologic
● Lipemic Sample
● Leukocytosis
● Hgb S and Hgb C

● HCT or Packed cell volume is the volume of

packed RBC that occupies a given volume of
blood.
● Reported as % or L/L
● Use EDTA or Heparin as anticoagulant
● Seal with clay at least 4 mm long
● Centrifuge for 5 mins between speed of 10,000
x g-15,000 x g
● Two methods

○ Macrohematocrit (uses wintrobe tube)

○ Microhematocrit (uses capillary tube)

- Male 40-54% (0.40-0.54 L/L)

- Female 35-49% (0.35-0.49 L/L)
- Newborn 53-65% (0.53-0.65 L/L)
- Child 30-43% (0.30-0.43 L/L)

Falsely Decrease ↓

• Incomplete sealing of tubes.

• Hemolysis
• Over Anticoagulated Blood

Falsely Increased ↑

• Reading including the buffy coat

• Inadequate centrifugation
• Abnormal RBC Morphology: Macrocytic Anemia,
Spherocytosis, Thalassemia, Hypochromic Anemia,
Sickle Cell Anemia