OUR LADY OF FATIMA UNIVERSITY

COLLEGE OF PHARMACY
Valenzuela. Quezon City. Antipolo. Pampanga. Cabanatuan. Laguna

BLOOD COLLECTION
Pharmacology Laboratory
At the end of the unit, the
student should have
1. observed proper
techniques in blood
collection
2. computed the
allowable volume of
blood to be withdrawn
from the test animals
In order to successfully complete Exercise 3,
please do the following:

1.Read: Introduction to Exercises 3 (Blood Collection)

2.Watch: Video links
3.Attend: Online Pre-laboratory Discussion via Zoom
4.Discuss: Join your classmates in a Group Discussion
5.Research: Work individually to answer worksheets
6. Quiz: Take online Quiz No. 2 (Topic: Methods of Drug
Administration and Blood collection)
 General
Considerations

Blood
collection
Blood Blood collection
collection NOT requiring
requiring anesthesia
anesthesia
VIDEO LINKS

Orbital sinus blood collection from rat

https://www.youtube.com/watch?v=xS25K1FmrFc

Cheek blood from mouse

https://www.youtube.com/watch?v=blTUVOB9yC8
 Blood Collection
GENERAL CONSIDERATIONS:
The area of injection or incision should
be cleaned with alcohol. “EXSANGUINATION”
Some procedures will require sedation
or anesthesia; others may be carried
out with a suitable restraint.
Upon completion, ensure good
hemostasis before returning the
animals to the cage.
 Blood Collection
Vasodilation
- using (+)-limonene will occur within 2-5
minutes and last for 10 minutes
(but after the procedure, apply 2% nitroglycerin ointment
to the affected site)
- immersing the tail in warm water for 5 to 10
seconds
- by warming the animal with a low-wattage
light bulb for 5 to 15 minutes prior to
venipuncture.
 Blood Collection
Species Circulating blood volume
(mL/kg BW)
Mouse 80
Rat 70

•10% of the circulating blood volume can

be taken at a 3-4 week interval
• At 24-hr interval, 1% of the circulating
blood volume may be withdrawn
 Blood Collection
Allowable volume of blood that may be
withdrawn from a 40-gram mouse

- in a 3 to 4-week interval?
(80 mL/kg x 0.04 kg x 0.10) = 0.32 mL
- in a 24-hr interval?
(80 mL/kg x 0.04 kg x 0.01) = 0.03 mL
 Blood Collection
NOT Requiring anesthesia

 Submandibular vein
 Saphenous vein
 Dorsal pedal vein
 Tail clipping
 Blood Collection
Through the submandibular area
• Animal lancet or small syringe needle may be used
to puncture the back of the cheek pouch

Orbital and
Submandibular veins
Puncture location
 Blood Collection
Saphenous venipuncture
• Shave the hair away from
the lateral and dorsal area
around the ankle joint
using a blade
• Small sterile syringe needle
is used to puncture the
skin
• Drop of blood formed at
the puncture site is
collected using a
hematocrit tube
 Blood Collection
Dorsal pedal venipuncture
 Warm the mouse and place
it in a restraining tube.
 With your thumb and first
finger, hold a hind foot
around the ankle. Your
thumb should be on top of
the foot. The medial dorsal
pedal vessel is found on the
top of the foot.
 Apply petroleum jelly to the
foot.
 Puncture the vein with a 23–
27 gauge needle.
 As drops of blood appear
collect them in a capillary
tube.
 Blood Collection
Tail-tipping/clipping
 For small-volume collection
 length of the tail that can
clipped :
less than 1 cm
 NOTE: No more than two
tail-tippings per animal
 Blood Collection
Retro-orbital sinus venipuncture
• For large-volume
collection
• Anesthesia is required
• Insert hematocrit tube or
pasteur pipet at the
medial canthus of the
eye
• Rotate tube in back of
orbit until blood flows
• Instill antibiotic eye
ointment after the
procedure
 Blood Collection
Requiring anesthesia
 Intracardiac puncture
 Posterior vena cava
 Orbital sinus
 Axillary vessels
 Blood Collection
Intracardiac puncture
Diaphragm

Last rib
Heart

Diaphragm

For large-volume collection Anesthesia is required

 Blood Collection
Frequent blood collection without replacement may
lead to short-term hypovolemic shock (longer -anemia)

% blood loss
10 Hemostatic cholinergic
mechanisms

15-20 Reduced cardiac output

and blood pressure

30-40 Hemorrhagic shock

 Euthanasia

 inducing painless death to laboratory animals

 done rapidly, humanely and away from the
public view
 reasons:

✓ sample collection
✓ post-mortem examination
✓ to free the animal from pain or distress
✓ to prevent animals from being “recycled”
 Euthanasia

Physical forms of euthanasia:

 Cervical dislocation
 Must be performed rapidly to achieve separation at
the cervical (neck) region and NOT the back.
 Decapitation

 Anesthetic
overdose
 CO2 chamber
 21

Thank you!
Any questions?

You can find me at:

smorencio@fatima.edu.ph

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