Protargol : silver impregnation of ciliates

Images made through a microscope. All subject types.

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Protos
Posts: 346
Joined: Tue Jan 07, 2014 4:01 am
Location: Lille, France

Protargol : silver impregnation of ciliates

Post by Protos »

I am getting back to this nice technique to identify ciliates using silver impregnation. There is space for improvement but it's getting interesting
Stylonychia sp
Image
Oxytricha sp. (TBC need more info on mid ventral cirri)
Image
Image
Last edited by Protos on Thu May 08, 2014 11:35 pm, edited 1 time in total.
Zeiss Axiophot, transmitted and Fluorescence
BK5000, Transmitted and CP
Wild M20

Beatsy
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Location: Malvern, UK

Post by Beatsy »

Yes, very interesting! Can you describe more about the technique you used ? Are the ciliates still alive in these images or does the staining protocol kill them? Cheers. Beats.

Protos
Posts: 346
Joined: Tue Jan 07, 2014 4:01 am
Location: Lille, France

Post by Protos »

In order to identify ciliates and for taxonomic evaluation, silver impregnation techniques are almost indispensable. The methods that are most frequently used are the dry and wet silver nitrate impregnation methods from Klein Chatton-Lwoff, the silver carbonate staining method from Fernandez-Galiano (which I use also see some of my posts) and the protargol impregnation methods by Dragesco, Deroux and Tuffrau. First discovered by the French school it is now improved by many including Foissner.
The methods I use here include about 30 steps which main parts are:
fixing ciliates with Boin
mounting them in glycerol albumin on a slide
Clarifying ciliates with K permanganate and oxalic acid treatment
staining with silver proteinate at 60°C
stopping impregnation with sodium hyposulfite
finally dehydrating and mounting in Balsam.

A slightly better view from belowImage
Zeiss Axiophot, transmitted and Fluorescence
BK5000, Transmitted and CP
Wild M20

Beatsy
Posts: 1840
Joined: Fri Jul 05, 2013 3:10 am
Location: Malvern, UK

Post by Beatsy »

Thanks for the info. Not a simple process then - congratulations on your excellent results. Particularly impressed that you end up with permanent mounts that are surprisingly well preserved. Very cool.

Bruce Taylor
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Location: Wakefield, Quebec / Ottawa, Ontario
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Post by Bruce Taylor »

Wonderful!

Do you have a commercial supplier for the protargol, or did you synthesize your own?
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Jean-marc
Posts: 274
Joined: Sat Oct 06, 2007 2:57 pm
Location: France

Post by Jean-marc »

Congratulation Protos, it is very good.

JM

Protos
Posts: 346
Joined: Tue Jan 07, 2014 4:01 am
Location: Lille, France

Post by Protos »

Bruce Taylor wrote:Wonderful!

Do you have a commercial supplier for the protargol, or did you synthesize your own?
I had some old protargol left, it works well at 0.4%
The protargol from Sigma works at 1% but not as strong
I will try to synthesize it according to the published recipe.

I will continue to use Silver carbonate for non-hypotrich ciliates; results are better and it is less time consuming !

Here comes a pic at 100x oil
Image
Zeiss Axiophot, transmitted and Fluorescence
BK5000, Transmitted and CP
Wild M20

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