Seeking a detailed procedure, because I gone through various article but did not get the step by step protocol. They just mention staining were done using borax carmine and further cercaria were mounted on glass slide with Canada balsam but my question is how to fix the invisible parasite and how to stain.
dear friend
its easy to perform, and I know how, but first you need to find the protocol and then do it based on it
Dear Rasool , thank you for your suggestion . I knew it is very easy procedure but thing is where to get it that step by step protocol . You said I know it , Can you give the link of that journal which mention the protocol very clearly. I gone through several article but I did not get that.
Please reply.
Dear Lalan Arya
It is really very easy to mouting larval trematodes. In our laboratory we use the following protocol
Kill the cercariae with water at 70 °C and add a fixative (formalin, ethanol 70 °or AFA).
Remove the fixative and put the stain and let overnight.
In the next day, remove the stain and dehydrates the larvae in alcohol series (ethanol 70°, 80°, 95°, absolute I, II, III, 15 minutes each). After alcohol III, add a clarifier (beechwood creosote or methyl salicilate) and mount in and in Canada balsam.
You can find other protocols in the following books: Schell (1965) How to Know the Trematodes. Dawes (1946) The Trematoda.
Best regards
Dear Hudson Pinto , Thank you very much for a valid answer. Actually I was wondering how to fix the cercaria on the slide, whether to first fixed on the slide or, stain it first and then put on the slide and mount it....!!!!!!
All the previous procedures must be done with the aid of dissecting microscope to observe the concentration of stain.
Also can use clove oil as clarifier. We use it in two steps: the first, mixed with absolute alcohol, and the second, with the clove oil pure. Then, follow with Canada balsam
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