MicrobeHunter.com Microscopy Forum

This is my second attempt at creating a "How to" video to demonstrate the proper technique for permanent slide preparation of whole mount insects. I created four ten minute videos. Technical difficulties with my IPhone prevented a smooth presentation but I think I got the important information across to the viewer. If you cut and paste the Youtube links below, the videos will play.

Video 1.
https://www.youtube.com/watch?v=jA1CDb9XjhE

Video 2.
https://www.youtube.com/watch?v=dUC2NiT8uUs

Video 3.
https://www.youtube.com/watch?v=u5gMCtMM3S0&t=131s

Video 4.
https://www.youtube.com/watch?v=hGRuI0kQb-U

Antoni

Excellent videos, very informative and interesting. Thanks for posting these.

Hi Antoni,
thank you for making and uploading these videos. I only found the time for a quick look but they seem to describe process and tools very well. I also like it that you didn't omit the safety aspect.

Bob

Thanks for these I had never even thought to attempt an exoskeleton so large so it is very interesting to see how it's done.

Very good series. Thanks for posting.

Great job. Very interesting and informative. Thanks for posting.

Interesting and informative series.
Concerning Xylene: Where did you get yours? The Xylene (Xylol) found in hardware stores is probably the same? This is commonly used in paint cleanup and doesn't really have any more warnings on the label as turpentine.
Concerning the dehydration method: Have you tried just dropping the specimen by itself in the jars of solution instead of on the slide? Seems like once they are dehydrated and ready to mount, they could be lifted out and arranged on the slide and mountant added? I've never done permanent insect slides but want to try some.

They will be too stiff to flatten to this degree after alcohol.

Thank you for making these videos available AntoniScott. You can read descriptions of how to do this, or have someone tell you, or see some photographs, but there's nothing like watching someone doing it in real time. Very much appreciated.
Tom W.

To Charles:
Your post did not mention your location, but in the USA ( where I live) you can purchase Xylene in large 3.8L (1 gallon) containers advertised as a "stronger than paint thinner" solvent from large hardware stores. As I stated in the video, Xylene is a known toxic solvent so I use an activated charcoal facemask with acrylo-nitrile gloves, outside where the ventilation is good, sometimes with a fan blowing away the xylene vapors from me.

Concerning your dehydration method question about just dropping the specimen into jars of solutions. The problem is that after processing a specimen in Potassium Hydroxide to soften the exoskelton and partially bleach the chitinous insect, it becomes very fragile. The chance of losing limbs, etc. is greatly increased with repeated handling and manipulating. Also, another member correctly stated that alcohol dehdration will stiffen an already fragile specimen. This will be further increased with xylene. Manipulation an insect at this point will almost guarantee that limbs will be lost.

I prefer to use my method to hold the insect fast between two slides separated with a paper shim at either end to prevent excessive crushing of the insect and to allow enough space for alcohol and xylene exchanges to take place.

Problem needing user attention:
I did not explain,or demonstrate the following suggestion in the previous videos (due time constraints) but alcohol and xylene exchanges do require sufficient space between the glass slides. When I move the glass slides containing the specimen from one container to the next ( i.e. from 90% alcohol to 100% alcohol or 100% alcohol to xylene #1) I usually place the end of the slide on an absorbent cloth of paper to "wick" the thin layer of solvents between the glass before transferring it to the next container. Although probably an excessive procedure on my part, once the slides have reached xylene, I have seven ( #1 - #7) exchanges (24 hrs per exchange) of xylene to ensure all of the fat and water is removed from the specimen.

Charles said: "Seems like once they are dehydrated and ready to mount, they could be lifted out and arranged on the slide and mountant added".

To Charles: This is possible (I used to do it this way) but risk of specimen limb loss, etc is too high. I developed my current technique through trial and error.

Antoni

very helpful, thank you

Great video series. Very informative. I'm going to have to implement some of these techniques.

On a side note, we seem to have been born under the same star. My other hobby is playing my Hauptwerk organ.

Amazing. I really enjoyed it. Thanks for the videos.

ps. It would be great if you make them "Save"-able. Currently, the save button is disabled.

To Blue:
I have to admit that I am unaware of the existance of a "save" button. If you could explain how to do that I would be glad to enable the save feature. Although I am being cynical, I can't imagine why a program would give the person the option to prevent someone from saving it.