I'd like some advice

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Greg Howald
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Joined: Tue Oct 20, 2020 6:44 am

I'd like some advice

#1 Post by Greg Howald » Fri Oct 01, 2021 3:17 pm

It has taken a long time for me to figure out how to set up and maintain an aquascape. Results are wonderful. There are hoardes of paramecium at the surface. I don't know what other things live a different levels. All of these things are fragile, mostly water and disintegrate when they die.
How can I make permanent slides? Do I need to heat set the specimens? Should I add dye while they are living?
I'd like some advice. I have some fairly nice equipment and many accessories but I'm still new to this. I seem to have to right stuff without the knowledge and experience required.
Thanks, Greg

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75RR
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Re: I'd like some advice

#2 Post by 75RR » Fri Oct 01, 2021 5:38 pm

.
No doubt some more knowledgeable members will chip in soon, but in the meantime you might want to look into Walter Dioni's articles.

I believe some of them cover your area of interest.

https://www.microscopy-uk.org.uk/mag/wd-articles.html
Zeiss Standard WL (somewhat fashion challenged) & Wild M8
Olympus E-P2 (Micro Four Thirds Camera)

Greg Howald
Posts: 1185
Joined: Tue Oct 20, 2020 6:44 am

Re: I'd like some advice

#3 Post by Greg Howald » Fri Oct 01, 2021 7:24 pm

Wow! That's a lot of info. Looks like I have some studying to do. Thanks.😃 Greg

TonyT
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Re: I'd like some advice

#4 Post by TonyT » Mon Oct 04, 2021 2:06 pm

A very difficult subject to make a permanent mount. First problem is to get the Paramecium to stick to the slide or coverslip. Standard practice is to mount them on a coverslip, but easier to handle when mounted on a slide. Mix equal parts of egg white and distilled water, shake well and filter. Apply a very thin smear to very clean slide (absolute alcohol with a drop of hydrochloric acid recommended for cleaning slides, dry slide with filter paper).
When almost dry add a drop of water containing Paramecium and spread as thin as possible. Gently heat the slide to coagulate the albumen (egg white).
The Paramecia should stick to the slide.
Next you have to 'fix' the specimens. Standard practice is to hold the slide upside down over a bottle of 1-2% Osmic Acid for 10 second; the fumes will fix the specimens (Osmic Acid is dangerous, likely impossible to obtain unless you are a recognized laboratory).
In the absence of Osmic Acid you could try adding a drop of Bouin's Fixative for about 5 mins. If no Bouin's you could try 99% Isopropoyl Alcohol.
Then need to stain the Paramecia with an alcoholic stain. Dehydrate, mount.
New Brunswick
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BramHuntingNematodes
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Re: I'd like some advice

#5 Post by BramHuntingNematodes » Mon Oct 04, 2021 4:48 pm

Do charged histology slides work with little animalcules
1942 Bausch and Lomb Series T Dynoptic, Custom Illumination

Greg Howald
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Re: I'd like some advice

#6 Post by Greg Howald » Mon Oct 04, 2021 5:25 pm

Thanks Toni. I have the stuff to try that. Much appreciated.
Greg

Greg Howald
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Joined: Tue Oct 20, 2020 6:44 am

Re: I'd like some advice

#7 Post by Greg Howald » Tue Oct 05, 2021 9:57 pm

To 75RR. Thanks for your help. I ordered the book and it arrived today. I will read it. Who knows? I might actually learn something if I try hard enough to get the information through my thick skull. I may have to drill a hole to pour it in.😆😆
Greg

To Toni.
I have tried your method and obtained some success.
I heat set the specimens by using my incubator. I undoubtedly need more practice but I am sure the method will work. Thanks. 😆😆
Greg

Greg Howald
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Joined: Tue Oct 20, 2020 6:44 am

Re: I'd like some advice

#8 Post by Greg Howald » Wed Oct 06, 2021 4:04 am

Today's results were fair using the egg white process.
I changed it up after that. I mixed clear Elmers glue with water, reducing the viscosity by about fifty percent, and painted each slide with it. I put the slide tray into the incubator at 105 degrees F. Until dry.
I removed the slides and placed one drop of pond water on each slide and returned the slides to the incubator for four hours. When dry I removed them and placed them two at a time upside down over a Petri dish containing 96 percent alcohol for three minutes. I dyed some with diluted alcohol stain and some with vital stains, and left them to dry. Those results were as good if not better than playing with egg whites or heat setting.
Tomorrow I will attempt to make really good permanent slides.

I also ordered the lactic acid for the GALA process given in the book and will try that when the acid arrives next week.
This has all gone from confusion to positive results in a very short time and I can't thank you folks enough.

I'm looking forward to dark field, oblique, rheinberg, and phase contrast.
Greg

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