Microscopy Lesson

Microscopy Lesson

Purpose: To observe fluke stages in saliva and urine with a
microscope.
Materials:
a. A low power microscope. High power is not needed. A
total of 100x magnification is satisfactory for the four
common flukes, Fasciolopsis, sheep liver fluke, human
liver fluke and pancreatic fluke.
b. Glass slides and coverslips.
c. A disposable eye dropper.

d. For sanitation purposes (wiping table tops, slides, microscope
and your hands) a 50% to 70% alcohol solution (not
rubbing alcohol!) is best. Dilute 95% grain alcohol 7 parts
alcohol plus 3 parts water. Vodka or 76% grain alcohol
can be used undiluted.
e. Formaldehyde, 20%. Formaldehyde 37% is commonly
available at pharmacies. Dilute this with equal parts of
filtered water to get 18½%, which is close enough to 20%,
for the purpose of “fixing” (killing) the specimens. Store in
a glass bottle in the garage, away from sunlight. Label.
Specimens that are fixed properly do not lose their life-like
appearance.
f Iodine solution. This is only useful for the urine specimens.
Lugol’s iodine and tincture of iodine are both useful. Ask a
pharmacist to prepare Lugol’s Iodine Solution for you, as
follows:
• 44 grams (1½ oz) iodine crystals
• 88 grams (3 oz) potassium iodide crystals
Dissolve both in 1 liter (quart) filtered water. This may
take a day of frequent shaking.
Method for saliva:
1. Pour the 20% formaldehyde into a small amber bottle or
other receptacle to a depth of about 1/8 inch. Keep tightly
closed.
2. The person to be tested is asked to salivate into the bottle
so the organisms are immediately “fixed” without undergoing
cooling first. The total volume should be about
double the original amount of formaldehyde used. Make a
mark on the container so the subject knows how much to
produce. The resultant concentration of formaldehyde will
be about 10%.
3. Shake the bottle a few times. Set it aside for 24 hours to
settle (less if testing is urgent).

4. With a dropper, draw up some of the bottom settlings. Put
one drop on a slide and apply a coverslip.
5. View under low power of microscope. Compare objects
you observe with specimens obtained on slides from biological
supply companies.
Note: Persons with HIV and moderate AIDS will show about
one to ten parasite stages per slide. It requires several hours of
searching. Persons with HIV and severe AIDS show 10 or more
fluke stages per slide; this makes the task of finding them much
easier. Persons with terminal untreated cancer have many more
fluke stages than relatively well persons.
Method for urine:
1. Prepare bottles of formaldehyde fixative ahead of time. Put
about ¼ to ½ inch of 20% formaldehyde in each. Keep
tightly closed.
2. Add freshly voided36 urine from cancer or HIV sufferers to
the formaldehyde in approximately equal amounts, resulting
in a 10% formaldehyde solution. Shake immediately. Let
settle several hours. The sediment has a higher number of
fluke stages. Cancer victims with cervical or prostate
cancer will show higher numbers of stages in urine than
other cancer types.
3. Staining the slide is optional. It helps to outline fluke stages
slightly. Prepare Lugol’s solution as described above.
Slides may be stained in either of these two ways:
• Put a drop of “fixed” urine on a slide. Add a drop of
50% Lugol’s (dilute 1:1 with filtered water). Apply
coverslip.

• Put a drop of “fixed” urine on a slide. Apply coverslip.
Add 1 to 3 drops of 50% Lugol’s to edge of
coverslip and allow it to seep in.
Note: persons who have been treated for cancer or HIV using
any of the known drugs may show only 1 to 2 fluke parasite
stages per drop of saliva or urine. For this reason, you may need
to search through 20 or more slides to find flukes. Very ill persons
may show up to 10 parasites per drop (slide).