Microscopic Staining for Blood Parasites

This six minute video demonstrates staining methods for microscopic diagnosis of malaria, including Field's (rapid) staining and Giemsa staining. This resource was developed by Charles Adjei Osei of the Kwame Nkrumah University of Science and Technology and Cary Engleberg of the University of Michigan. It is part of a larger learning module about laboratory methods for clinical microbiology. The full learning module, editable animation, and video transcript are available at http://open.umich.edu/education/med/oernetwork/med/microbiology/clinical-microbio-lab/2009. Copyright 2009-2010, Kwame Nkrumah University of Science and Technology and Cary Engleberg. This is licensed under a Creative Commons Attribution Noncommercial 3.0 License http://creativecommons.org/licenses/by-nc/3.0/.

(English captions by Andrea Matsumoto, University of Michigan.)

Take venous blood sample into a sequestrated
air bottle mixed very well.

Take a drop onto a microscope slide.

We can also as well take a finger prick from
the third or ring finger.

Using a microscope slide as a spreader, spread
the blood, drop of blood, into a thin form.

Air-dry the smear, the glass smear on a draining
rack.

Prepare a thick smear by putting blood from
a sequestrated bottle onto the microscope

slide or as well you can use blood from a
finger prick.

Using a microscope slide spread the blood
out.

Allow it to air dry on the draining rack or
in the incubator.

Field's Rapid Staining for Malaria:

Stain the thick smear in a field stain A for
five seconds.

And this dehemoglobinizes the red blood cells.

Wash under the tap water or buffered water.

Put it in field stain B for five seconds.

After which you will wash under the tap water
or with buffered water.

Air-dry it on the draining rack.

Fix the thin smear in methanol for two seconds.

Allow it to air dry and put it in field stain
B for six seconds.

After which you wash under tap water or buffered
water.

Put it in field stain A for six seconds.

After which you wash under the tap water to
get rid of the excess stain and this stain

is called river staining and this preserve
the red blood cells.

The Giemsa Stain:

Stain the thick smear in one in ten dilution
of Giemsa for five to fifteen minutes.

Wash the excess stain under tap water or buffered
water.

The thin smear is fixed in methanol for two
seconds after which it is put in the Giemsa

one in ten dilution for five to fifteen minutes.

Wash off the excess stain under tap water
or using buffered water and air dry it on

draining rack.

Examine the thin slides under times hundred
(100x) using oil immersion.

It is better to see for the thick film the
parasites and the white blood cells.

The parasites will look like a ring form or
dots, which appear in pairs.

But in a thin film the parasites are found
inside the red blood cell for which we can

use to determine the species of the parasites.