Information about tapeworms staining methods

[Razielox]

Information about tapeworms staining methods

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Post by Razielox » Mon Oct 23, 2006 8:43 pm

Do you guys know some sites which have some information about staining of tapeworms ?

[Dr.Stein]

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Post by Dr.Stein » Thu Oct 26, 2006 8:58 am

I don't know any sites but according to my experience, I applied HE staining to study the morfology and anatomy of tapeworms and it works well. What do you want to study of your tapeworm?

[Razielox]

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Post by Razielox » Mon Oct 30, 2006 11:05 am

if you don't know any sites wchich may contain some staining methods of parasites, it's a pity.
We have some old methods with HE staining in our lab, but these are some old methods without details - i mean there is entire method step by step however we don't have for example proper time schedule regarding how long the segment of tapeworm should be hold in alcohol to be dehydrated.
These our methods were written by one doctor who did many
attempts when he was working here with all these parasites and he made many nice preparations for students. There is lack of descriptions on the internet about stainig methods of tapeworms, but maybe YOU have some informations and you can share with me.
Sometimes the tapeworms are very nice after staining and some time there are very bad - and not every time we know why...
And because of lack of material in these days it could be good to know how to do it properly ( staining ), bacuse we don't want to waste our precious tapeworms

[Dr.Stein]

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Post by Dr.Stein » Tue Oct 31, 2006 6:14 am

As I only wanted to watch the anatomy of tapeworm at that time without special treatment, thus applying HE staining is enough and the result is not bad at all. I follow the basic protocol but did modification here and there because, according to my experience, by applying the original steps will destroy the tissues. For instance, I couldn't use formaldehyde to fix it because the tissue will be fragile. And the clearing process with toluene couldn't be apllied overnight, but only few hours til the tissue becomes transparent. Then the application of dye must be modified as well, because I don't like Eosin too much for my preparation. I prefer more hematoxylin. The cuticula of tapeworm absorbs Eosin too much so I judt did some seconds on Eosin and longer in hematoxylin. About the lenght of the tapeworm for the sample, it depends on what you want to see. When I want to watch the visceral, I just need half of the segmen. When I wanted to watch how the segmen is arranged to another, I cut not right on the junction but some milimeters right and left from it. This HE method stains visceral very well, especially the eggs

[Razielox]

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Post by Razielox » Tue Oct 31, 2006 8:13 am

Well - you seems to know a lot in this subject.
Do you have maybe some schedules, or some descriptions in your home or somewhere which may show how to stain one segment of tapeworm with HE?
We would like to see excretive ducts and reproductive ducts.
It would be fine to have whole staining procedure which describes it from begining to the end of staining with all the details.
Do you have some ? I'd be grateful.

[Dr.Stein]

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Post by Dr.Stein » Tue Oct 31, 2006 9:10 am

Well, basically I follow McManus' HE staining method. Then, all you need to do is modifying it, e.g. the duration of the staining, to obtain your favorite staining.

1. Deparaffination - xylene about 30 minutes
2. Hydration - ethanol (staged) to water/distilled water (dip just a while)
3. Staining - Ehrlich Hematoxylin about 1 minute
4. Rinse the dye with flowing tap water about 10 minutes or til clean
5. Dehydration - ethanol (staged) to ethanol 70 % (dip just a while)
6. Staining - Eosin-Y about 30 seconds
7. Dehydration - ethanol (staged) to ethanol absolute or 96 % (dip just a while)
8. Mounting - canada balsam

Feel free to ask me, I try to explain as far as I can

[sachin]

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Post by sachin » Tue Oct 31, 2006 9:59 am

We use same method for permanent mounting.

This is ultimate method for any organism to make permanant slide in small lab level.

Alcohol----10%-30%-50%-70%-90% each for 20 sec.
stain--- methyl red / eosine 20 sec
alcohol-----absolute 20 sec

mount with DPX.

Observe.

[Razielox]

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Post by Razielox » Tue Oct 31, 2006 11:11 am

One question for you Super Member:

1. Do you use paraffin to preserve your segments of tapeworm ?

We staining our segments with Karminum rubrum with natrium boricum.
I think that your answers and descriptions will be helpful for us - thank you.
If we have some more questions i will write them here.

[Razielox]

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Post by Razielox » Tue Oct 31, 2006 12:18 pm

by "Super Member" i meant Dr.Stein

[Razielox]

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Post by Razielox » Tue Oct 31, 2006 12:26 pm

And one more question to Dr. Stein :

Can dryness of segments may be the cause for quality of vision after staining?
I mean if the segments were dry long time before they got to us, could this can be the cause for very bad quality, or total failure ?

[Dr.Stein]

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Post by Dr.Stein » Wed Nov 01, 2006 4:04 am

Answers for our Super Tapeworm Lover

1. Yes, simply I used paraffin to block the segmen. Or you can use paraplast to have a better preparation. At that moment I was lack of paraplast so I just applied paraffin and it worked well.

2. Dryness of specimen is fatal!!! Also let the specimen too long in open air after taken from the body before processing is a great loss! You should soak them into fixative as soon as possible after they are taken. You can apply ethanol 70 % for this tapeworm if you want to preserve in long time, never use formaldehyde. You can use 4 % formaldehyde for short preservation only or your tapeworm will be fragile when you process it by then.

Good luck