BLOOD COLLECTION

CEPHALIC VENIPUNCTURE

Purpose

To obtain a sample of venous blood for analysis

Indications

Collection of a blood sample for clinical pathology tests

Contraindications and concerns

Proper restraint is important to prevent excessive trauma to the vein, resulting in hematoma
formation.

Complications

1. Hemorrhage.

2. Subcutaneous hematoma formation.

Special anatomy

Cephalic vein: The right and left cephalic veins are superficial veins that lie on the anterior surface
of the forelimb, making them very accessible for venipuncture.

Equipment

• 22- to 20-gauge, 1-inch needle

• Syringe
• 70% alcohol
Restraint

1. Place the animal in a sitting position or in sternal recumbency on a table or (for large dogs) on
the floor.

2. The holder should stand on the side opposite the leg to be used and should use one arm to
restrain the animal’s head by encircling the neck and turning the muzzle away from the leg to be
used. The holder should use the other arm to extend the animal’s front leg by holding the elbow
and pushing the leg forward.

Technique

1. Laterally roll and compress the cephalic vein.

Using the thumb of the hand holding the leg, the cephalic vein is rolled laterally and compressed so that it becomes
distended with blood.

2. If the vein cannot be seen or palpated, clip hair from a small area over the dorsal forearm, and
apply alcohol.

3. The venipuncturist should grasp the paw to keep the leg extended. He or she should identify the
distended cephalic vein and place the thumb alongside the vein to stabilize it during venipuncture.

4. Insert the needle, bevel upward, at a 20- to 30-degree angle to the vein. Once the tip of the
needle is in the vein, apply suction to collect the sample.

5. Once the sample is collected, have the holder release the pressure on the vein. Halt suction and
withdraw the needle from the vein. Place gentle pressure on the venipuncture site, and hold for
approximately 60 seconds.
 BLOOD SMEAR PREPARATION

Type of procedure

Diagnostic sample collection

Procedure explanation and related physiology

Examination of a blood smear is an integral part of a complete blood cell count. When examined by
a trained technologist or pathologist, a blood smear can provide a large amount of information
about RBCs, WBCs, and platelets. Preparation of a smear at the time of sample collection can
provide a snapshot view of the patient’s hematology status, minimizing the effects of sample
deterioration, such as changes in leukocyte morphology and decreased platelet numbers. Hence,
even samples sent to a diagnostic laboratory should include an air-dried smear of fresh blood.

Poorly prepared smears can either mask abnormalities or result in confusing artifacts. RBC and
WBC morphologic features can be obscured by extremely thick smears, whereas changes in cell
distribution on the smear due to uneven streaking can lead to serious errors in the differential
count. Mastering the technique for preparing a good-quality blood smear takes some practice.

Indications

• When transport to a diagnostic laboratory delays sample processing

• An essential part of an in-house CBC CONTRAINDICATIONS Blood is clotted—results may be
erroneous.

Body systems assessed

Hemic, lymphatic, and immune

Preparation

• Clean the venipuncture site with alcohol.

• Perform venipuncture and transfer blood into a tube containing EDTA (lavender top).

Equipment or supplies

• Clean glass slides

• Microhematocrit tubes (an optional way to transfer blood to slide)
• Wooden applicator sticks

Technique

• Swirl a pair of applicator sticks in blood to check for blood clots.

• Place a drop of well-mixed blood at 1 end of a glass slide.
• Place a second glass slide at a 45◦ angle to the first slide, with the blood droplet positioned
between the 2 slides (Figure 1). Avoid pressing down on the spreader slide.
• Smear thickness can be adjusted by changing the angle between the 2 slides. With samples
from severely anemic animals, widening the angle between slides can help avoid making too
thin a smear.
• Slide the top slide toward the drop of blood.
• When the spreader slide contacts the droplet, pause briefly, allowing blood to spread along the
juncture between the 2 slides.
 • Move the spreader slide away from the drop of blood in a smooth, steady motion. The
spreader slide should not be lifted until it has reached the far end of the lower slide.
• Allow the smear to air-dry and stain with some type of Romanovsky-type stain (e. g., Diff-Quick,
Hema III, Giemsa, Wright stain).

Sample handling

• Air-dried blood smears can be stored several days at room temperature.

• Once fixed and stained, smears are stable for months to years if protected from light.
• A coverslip is needed for crisp resolution at 40-fold magnification. A temporary coverslip can be
attached with a drop of immersion oil. Commercial mounting media can be used to attach a
coverslip permanently, which also protects smears from scratches, dust, etc.

Normal findings or range

• A bullet-shaped blood smear that extends about one-third to two-thirds the length of the glass
slide.
• The smear has a zone just behind the feathered edge in which RBCs are closely opposed to
each other with little overlap (monolayer).
Conditions that may interfere with performing the procedure

Marked polythemia may prevent preparing a smear with a monolayer and cause distorted
leukocytes.

Procedure techniques or handling that may alter results

• Using poorly mixed blood can result in a thin smear suggesting anemia.
• Using clotted blood can result in artificially decreased platelet numbers.
• Platelets are more likely to be clumped in old blood (>6 h).
• Leukocyte morphologic features can be altered in old blood.
• A thick, squared-off feathered edge or the lack of a feathered edge can prevent recognition of
platelet clumps or abnormal cells.
• Exposure of smears to formalin fumes produces a thick blue-green background and prevents
accurate leukocyte identification.

Clinical perspective

• Even precleaned slides can have glass chips on their surface, which should be wiped off.
• A smear of freshly collected blood may be needed to identify RBC parasites such as
Mycoplasma hemofelis.
• A smear of freshly collected blood provides the most accurate platelet estimate.
• Capillary blood (e. g., from ear prick) can be helpful for finding RBC parasites such as Babesia
sp.