BIO03-Biotechniques

2nd AY 2015-2016 1
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Exercise # 4
Preservation of Nematode eggs

I. Objectives:
At the end of this activity, the student should be able to:
1. provide a prepared slide using egg fixation
2. preserve the nematode eggs efficiently
II. Materials:
Microscope slides
Cover slips
Cotton
Coplin jar
Xylene

Canada balsam/Histomount
methanol (100%)/ petroleum ether
90% ethanol
Alcohol, 70%
Absolute alcohol

III. Procedure:
1. Transfer a drop of well-mixed feces preserved in formalin to a slide
and spread it uniformly (size of a dime)
2. Add methanol/petroleum ether (100%) and dry
3. Leave the slide in a Coplin jar with 70% ethanol for 5 to 10 minutes,
transfer to 90% ethanol (3-5minutes) and finally to absolute alcohol
(5-10minutes). The time frames can vary; longer duration leads to
better dehydration
4. Transfer the slide from absolute alcohol to xylene (3-5minutes)
5. Overlay with 2-3 drops of Canada Balsam/histomount, add a cover
slip, leave the slide in the incubator for 24-48 hours for drying.
IV. RESULTS. Please attach picture of the prepared slide and the
specimen under the microscope