Blood smear technique for veterinarians

Below is a step-by-step guide for veterinarians produced by the Department of Agriculture and Food, Western Australia for
performing blood smears for diagnostic use.

Examination of blood smears is an integral part of haematological testing. Blood smears are useful for examining cell morphology
and identifying blood-borne parasites. Remember to include a detailed history and description of lesions on the DAFWA
Diagnostic Laboratory Services (DDLS) submission form.

For more information on blood smears, contact DDLS on +61 0(8) 9368 3351.

See the links on this page or email DDLS@agric.wa.gov.au for laminated copies of other diagnostic reference guides A visual guide
to a ruminant animal post-mortem, Basic diagnostic sampling protocol – livestock, Brain removal techniques and A visual guide to a
chicken necropsy.

Perform blood smears when you suspect:

blood-borne protozoal or mycoplasmal conditions
haemolytic conditions
anaemia
haematologic neoplasia.
Equipment required for performing blood smears:
microscope slides with frosted ends for writing on
spreader slide – use a specific spreader slide with bevelled edges, or a second clean microscope slide
capillary (microhaematocrit) tubes
pencil to label the slides
blood collected in an EDTA tube
pot of sterile water to clean the spreader slide between uses.

Top tips for making good smears:

Making great blood films takes practice; don’t be discouraged by less than perfect smears. Submit all blood smears, perfect or
otherwise, as areas of the film may be suitable for examination.

Collect blood in an EDTA tube and make the smears when back at the clinic.
Use clean, high-quality microscope slides.
Aim for a blood droplet size of 4mm diameter.
Optimise spreading speed for length and a good feathered edge.
Hold the spreader slide at 30-40 degrees to achieve optimal smear length.
Maintain even contact throughout the spreading motion.

Step 1
 Fill a capillary tube three-quarters full with blood.
Hold the capillary tube vertically over the slide.
Allow 1 drop of blood to form on the end of the tube.

Step 2
 Place the tube onto the slide about 0.5cm from the frosted area.
Leave a drop of blood about 4mm in diameter on the slide.

Step 3
 Hold the spreader slide or second microscope slide at a 30-40 degree angle at the end of the slide (i.e. in front of the blood
droplet).
Ensure the short edge of the spreader slide is in even contact with the lower slide.
Pin the lower slide to prevent it from moving.

Step 4
 Using a smooth motion, draw the spreader slide back through the entire drop of blood.
Allow the blood to spread evenly along the edge of the spreader slide.

Step 5
 Push the spreader slide forward along the length of the lower slide.

Step 6
 Maintain a constant smooth motion, angle and even contact.
Note: blood is being dragged behind the spreader slide, not in front of the slide.

Step 7
 An optimal smear is three-quarters the length of the slide and has a feathered edge.
Leave the slide to air dry and make more smears if required.
Pack smears individually into slide holders.

Unstained (left) versus stained smears

Do not fix or stain the smears. The DAFWA Diagnostic Laboratory Services (DDLS) will do this under controlled conditions to
optimise the staining process.

Troubleshooting blood smear errors

Blood smears - common errors

The blood droplet on the left is too big and will result in a thicker smear. The blood droplet on the right is the preferred size.

Smear technique - left to right:

slide 1 - perfect smear

 slide 2 - smear technique interrupted in
Reference: https://ahdc.vet.cornell.edu/Sects/ClinPath/sample/test/hema.cfm middle
slide 3 - smear was skewed
Problem Solutions slide 4 - blood droplet too thick
slide 5 - smear too short.
Use a larger droplet of blood.
Decrease the angle of the spreader slide.
Short smear Infectious diseases in
Decrease the speed of the spreader slide.
Australia diagnosed by blood
smear
Use a smaller drop of blood.
Long smear/no Increase the angle of the spreader slide.
feathered edge Increase the speed of the spreader slide.

Bovine erythrocytes infected with protozoal

Use smaller drop of blood.
Decrease the angle of the spreader slide.
Thick smear
Increase the speed of the spreader slide.
Use a larger drop of blood.
Increase the angle of the spreader slide.
Thin smear
Decrease the speed of the spreader slide.
parasites from the Theileria orientalis
group. The parasite is known as a
piroplasm when it is within an erythrocyte.
Piroplasms appear in erythrocytes from
day 10 post-infection. Naïve, young,
pregnant or immunecompromised animals
may develop severe anaemia and
mortalities can be as high as 30% in a herd.

Maintain even contact and a smooth motion.

Increase the speed of the spreader slide.
Smear has waves and Haemaphysalis longicornis (common bush
Relax the wrist, reduce downward pressure on the
ridges tick)
spreader slide.

Ticks are vectors for many protozoal and

mycoplasmal parasites. For Theileria spp.,
transmission in the tick is known to be
Disease Species Transmission Disease status
transstadial or life stage-to-life stage. A
larva or nymph stage tick transmits the
endemic-northern
Anaplasmosis cattle tick-borne parasite to the next animal it feeds on.
Australia
Trans-ovarial transmission (transmission
from infected females to their larvae) does
endemic-northern
Babesiosis cattle tick-borne not occur. Control of ticks and good
Australia
sanitation when using needles and surgical
iatrogenic, blood- equipment minimises the introduction of
Mycoplasma ovis sucking insects these infectious parasites into naïve
sheep endemic populations.
(eperythrozoonosis) (midges, mosquitoes,
flies)

endemic in eastern
Bovine anaemia due to
Australia - first All Page Links
Theileria orientalis group cattle tick-borne
diagnosed in WA in [1] mailto:DDLS@agric.wa.gov.au
(BATOG)
2013 [2]

https://www.agric.wa.gov.au/sites/gateway/files/Equipment_0.jpg
[3] https://www.agric.wa.gov.au/sites/gateway/files/Step%201.jpg
[4] https://www.agric.wa.gov.au/sites/gateway/files/Step%202.jpg
[5] https://www.agric.wa.gov.au/sites/gateway/files/Step%203.jpg
[6] https://www.agric.wa.gov.au/sites/gateway/files/Step%204.jpg
[7] https://www.agric.wa.gov.au/sites/gateway/files/Step%205.jpg
[8] https://www.agric.wa.gov.au/sites/gateway/files/Step%206.jpg
[9] https://www.agric.wa.gov.au/sites/gateway/files/Step%207.jpg
[10] https://www.agric.wa.gov.au/sites/gateway/files/styles/original/public/stained%20vs%20unstained%20slides_1.jpg?
itok=m_3qseBp
[11] https://ahdc.vet.cornell.edu/Sects/ClinPath/sample/test/hema.cfm
[12] https://www.agric.wa.gov.au/sites/gateway/files/styles/original/public/blood%20smear-common%20errors%201.jpg?
itok=Fuz7AUpy
[13] https://www.agric.wa.gov.au/sites/gateway/files/styles/original/public/blood%20smear-common%20errors%202_0.jpg?
itok=KOgeZ5av
[14] https://www.agric.wa.gov.au/sites/gateway/files/styles/original/public/cytology%20slide-bovine%20theileria.jpg?
itok=2e3nMfYd
[15]
https://www.agric.wa.gov.au/sites/gateway/files/styles/original/public/Blood%20smear%20posterprint_Page_2_Image_0004_3.jpg?
itok=RQVAHbfk
[16] https://www.agric.wa.gov.au/livestock-biosecurity/chicken-necropsy-guide
[17] https://www.agric.wa.gov.au/livestock-biosecurity/ruminant-animal-post-mortem-guide
[18] https://www.agric.wa.gov.au/livestock-biosecurity/newcastle-disease-sample-collection-western-australia
[19] https://www.agric.wa.gov.au/livestock-biosecurity/sampling-and-post-mortem-resources-veterinarians
[20] https://www.agric.wa.gov.au/livestock-biosecurity/sampling-procedure-toxic-algae
[21] https://www.agric.wa.gov.au/livestock-biosecurity/brain-removal-techniques-livestock
[22] https://www.agric.wa.gov.au/sites/gateway/files/Master%20your%20blood%20smear%20technique.pdf

Source URL: https://www.agric.wa.gov.au/livestock-biosecurity/blood-smear-technique-veterinarians

This print version was generated at 8:44pm on the 21st of May, 2020.
The original document was last revised at Tue, 02/05/2017 - 4:26pm

Important disclaimer
The Chief Executive Officer of the Department of Primary Industries and Regional Development and the State of Western Australia
accept no liability whatsoever by reason of negligence or otherwise arising from the use or release of this information or any part of
it.