FORMALIN-ETHER

SEDIMENTATION
Zion Michelle J. Huete
BSMT II - M24
 Topic Outline

01 02 03
Introduction Definition Pros and Cons

04 05
Materials Procedure
CONCENTRATION PROCEDURES
• Concentration procedures involve isolating parasites from
fecal debris and enhancing the likelihood of detecting
parasitic organisms, particularly when they are present in
low numbers.
• These procedures are categorized into flotation and
sedimentation techniques.
 SEDIMENTATION TECHNIQUES
• Use solutions of lower specific gravity than
the parasitic organisms, thus concentrating
the latter in the sediment. Sedimentation
techniques are recommended for general
diagnostic laboratories because they are
easier to perform and less prone to technical
errors.
⚬ Formalin Ether Concentration Technique
⚬ Acid Ether Concentration Technique
⚬ Gravity Sedimentation
⚬ Centrifugal Sedimentation
⚬ MIF Sedimentation
FLOTATION TECHNIQUES
• Use solutions which have higher
specific gravity than the organisms to
be floated so that the organisms rise to
the top and the debris sinks to the
COMPARISON bottom.
⚬ Modified Zinc Sulfate Concentration
⚬ Brine Flotation Technique
⚬ Sheather’s Sugar Flotation Technique
 FORMALIN-ETHER SEDIMENTATION

Recommended as being the

Allows the broadest range of
easiest to perform and the least
Specimen can be fresh or fixed parasitic elements
subject to technical error
stool
 FORMALIN-ETHER SEDIMENTATION
Efficient for recovering most Concentrate specimens
helminth eggs, larvae, and preserved in formalin (10%)
protozoan cysts
Ether can dissolve neutral fats in
the stool
Sediments can be stored for a
long time • Ether is flammable,
replaced by ethyl acetate
Parasite morphology is better
preserved in formalin
ETHYL ACETATE
• more efficient in recovery of
cestode eggs and Giardia
cysts, however, not as
efficient in the extraction of
fat or mucoidal material from
the stool
ADVANTAGES 01 Recovers helminth eggs, larvae and
protozoan cysts
****ADVANTA
GES****
Used to concentrate formalin-
02
preserved fecal specimens

Specimens from FECT can be stored for a

03 long period of titme

More efficient in the recovery of cestode

04 eggs and Giardia cysts
 Ether is explosive and flammable
DISADVANTA
01
(ethyl acetate may be used instead)
GES*DISADVA
NTAGES*
not recommended for eggs of Fasciola spp.
02
and larvae of Strongyloides stercoralis.

Not that efficient in extraction of fat or

03
mucoidal material from stool
 MATERIALS

APPLICATOR STICKS BEAKER CONICAL TUBE

CENTRIFUGE FUNNEL GAUZE

SPECIMEN FORMALIN ETHYL ACETATE

MISCROSCOPE SLIDE AND COVER

DISTILLED WATER SLIP MICROSCOPE

Transfer 2 to 5 g (2 to 5 mL if
diarrheic) of feces to a paper
cup or beaker. The more
fibrous the stool, the more
specimen you will have to use
(up to 5 g)
STEP 1
STEP 6
Break up the sediment in the stool
sample. Add 10% formalin to fill half
of the tube and mix well. Let the
suspension stand for at least 5 minutes
for proper fixation. Optionally, stopper
the formalin-feces mixture for later
use.
STEP BY STEP PROCESS
Pour through 1 to 2 layers of wet
Add approximately 3 mL of gauze into a 15-mL conical tube.
water and comminute to a Leave the coarse debris into the cup
uniform suspension. by crimping the lip and pouring the
then add another 10 mL suspension through the slit formed.
water to the fecal suspension
and mix.
STEP 3
STEP 2
STEP 8
STEP 7
Centrifuge at 1500 rpm for 1 minute to
Add ethyl acetate to half
create three layers: formalin, fecal debris
fill the remaining space in
plug, and ether. Loosen the plug with an
the tube. Mix vigorously
applicator stick, discard everything except
for a full minute. Perform
the sediment. Clean tube walls with a cotton
this step away from any
swab to prevent debris from settling into the
fire.
sediment.
Fill the tube and centrifuge at
2000 to 2500 rpm for 1 minute.
and Carefully pour off the
supernatant. There should be
about 0.5 to 1.0 mL of
sediment in the tube.
STEP 4
STEP 9
Mix the remaining sediment with
drained fluid from the tube sides.
Adjust suspension density with water
if needed. Transfer a small drop to
iodine solution for protozoan cyst
examination. Cover with slip and
examine.
Break up the sediment and resuspend
with fresh water. Centrifuge the tube
and decant as before. Repeat this step
if the supernatant fluid is not
relatively clear.
STEP 5
STEP 10
Examination of the specimen can be delayed.
Add 1 or 2 mL of 10% formalin to the sediment
and stopper the tube. Formalized sediments can
be kept indefinitely. Remove the excess
formalin before making the mounts to prevent
diluting the concentration.
THANK
YOU