🔬 059 - How to make a PERMANENT SLIDE for Microscopy

Making a dry mounted slide and one with mounting medium. Making a spacer ring with a slide ringing table.
Ringing table from Brunel microscopes: www.brunelmicroscopessecure.co...
Here I show you how to prepare a wing of a wasp. The mounting medium used is Euparal, which is common for mounting insect specimens. In this video we will have a look at how the differently mounted specimens appear under the microscope.
🖂 NEWSLETTER - www.microbehunter.com/newslett...
🎈 BECOME A PATRON - / microbehunter
👜 MICROSCOPY SHOP - www.microbehunter.com/shop/
💻 WEBSITE - www.microbehunter.com
RECOMMENDED MICROSCOPY PRODUCTS (Affiliate links)
Swift SW380T: amzn.to/2UkTpXb
Swift SW350T: amzn.to/3BdSqbV
Swift 150: amzn.to/3ikisBC
Slides and Cover glasses: amzn.to/2VOCa0J
Slide preparation kit: amzn.to/3z9Aqxv
#microbehunter #amateurmicroscopy #microscope

[@usernamemykel]

Nice choice of eyeglasses, Oliver. To summarize: Bugs - dehydrate in alcohol, mount in Euparal. Liquid specimens - do not dehydrate, mount in Glycerin How about dehydrated bugs mounted in "New Skin"/"Liquid Bandage" or nail polish if Euparal isn't available? Many thanks!

[@Nextsession271]

I just bought my grandkids a microscope. Thankn You for simplifying the process for me. I'm a newbie at this. Great Video!👊🏿👏🏾👏🏾👏🏾👍🏿

[@PrinceZee]

Great channel man. Keep going.

[@KillerTr0nn]

I just wanted to thank you for these videos and channel. I always wanted to do this and your channel gave me the push that I needed. I got myself a 🔬 form the ones you recommend in Amazon. Super excited to start making permanent slides for my collection. Again, thank you very much. 😁

[@PolYYerP]

Amazing work and very informative. Keep on going!

[@christiandietachmair334]

Thanks for your great work !

[@DopravniPoradce]

Thank you very much for your videos. I just bought my first microscope and it's very useful to get some tips.

[@parapresumirhayquesufrir8736]

Great video glad I found this channel

[@ms.scienceandreasonlover]

Thank you this video helps a lot

[@f3derico2007]

Very informative

[@lemonpepperdry5818]

Thank you for this information.

[@kulsoomshah9225]

Amazing ✌️

[@sarahasamoah7475]

Woooow you're s great teacheerrr

[@shaheenmajeedwani1969]

Nyc work

[@Thecarverman]

Hello microbe hunter I was wondering if you could do a video on how to make a permanent slide of paramecium pleas I can’t find anything anywhere on this i thought you may be the one to show me and a lot of others on how to do this thank you so much your videos are extremely helpful and I hope all is good with you during these tough times with covid-19 🦠

[@DopravniPoradce]

I looked up Euparal and as far as I understand that you use a drop at a time, it's definitely not cheap. Equivalent of 91 USD for 100 ml in a Czech shop...

[@lavrentii]

I know that for some organisms, you can dehydrate them gradually, by using sequentially more concentrated alcohol. What's your experience with this?

[@Microbehunter]

Alcohol series works with some specimens. The step-wise dehydration is done with animal tissue in histology. Some water organisms (Paramecium etc) are alcohol sensitive and the alcohol will break the cell membrane, so an alcohol series won't work. As a consequence, they pop open and run out. Other organisms like plants will lose their pigments (chlorophyll) when in alcohol, even when dedydrating them in an alcohol series. Insects have a hard exoskeleton and this is more resistant to shrinking. I read somewhere that addition of acetic acid counter-acts the swelling somehow. I did not try this, however.

[@nokiot9]

If I don’t have a spinny table like that is there a good way to make the spacer? Do the like sell stick on ones? Because I’ve been just using those indented slides that have the little gouge dug out of them.

[@darwinmendoza2541]

More please

[@DSScully]

Do you have a video that discusses clearing samples before mounting?

[@DSScully]

I'm specifically interested in clearing microscopic insects ( thrips, psocopids, aphids, whiteflies)

[@raikj1078]

2:08 THE the Wasp

[@PaulAJohnston1963]

Love the video. Question How many slides do you think you can do with 50ml Euparal please?

[@Microbehunter]

I would have to do a bit of math here becasue I did not measure this myself. Let's assume that we use a larger 20mmx20mm cover glass and that the medium is 0.1mm thick. Per slide we would use 20*20*0.1 microliters = 40 ul. We then divide 50000ul/40ul and we would get 1250 slides. Maybe a bit less because some specimens are thicker and require more mounting medium.

[@PaulAJohnston1963]

@@Microbehunter Many thanks for the extremely quick response. I looked at Euparal found it at www.agarscientific.com/mountants-euparal and wondered how much to buy, do you recommend getting the thinner too?

[@Microbehunter]

I always dilute it with a few drops of ethyl alcohol, but if you keep the bottle closed well, then the Euparal will remain liquid for a long time. I have not had the need for the thinner, but some people who make slides that last a long time apparently also put the specimen for a few days into the thinner to make sure that it's completely soaked through with the same solvent. Never done that though. If thinner is cheap, you might as well get it. Maybe also helps remove excess solidified Euparal. Just a general warning, though. Don't put Euparal mounted slides directly under the microscope. Always wait until dry. Risk of getting the objective contaminated with it. Talking from experience here........

[@max-7404]

Is it okay if i put a square cover glass on the circle nail varnish ??

[@shoppersplace]

Sure, but edges may break

[@nikitatosur2547]

Sir I want to prepare thrips mount how to do it

[@AliRaza-eb3vf]

wasal di slide wah wah #saraikilearning

[@backyardmicrobiology]

i dont have euparal so i use glue..it dont work nice..is there anything to replace euparal for mount speciemen?

[@Microbehunter]

Micscape magazine has a comprehensive article of various mounting media: www.microscopy-uk.org.uk/mag/artjun04/wdmtmedia.html

[@thomasschwarz1973]

Here are some recipes www.aeisner.de/

[@tipswithreza1045]

Good effort #saraikiLearning

[@martyisabeliever]

I am in the USA and cannot find EUPARAL??? Not even on Amazon? 4/2019

[@THATHATSALLFOLKS]

AMAZOM HAS IT NOW

[@gulpgulpgulpgulp69]

where did you get the slide turntable you used to put the nail polish on the slide?

[@Microbehunter]

Ringing table from Brunel microscopes: www.brunelmicroscopessecure.co.uk/acatalog/universal.html

[@ErfanColorectal]

I have a problem about the mounting medium.my father bought me a introductory microscope some days ago.Now I want to make some permanent slides.But the mounting medium isn't available here. Can I use glycerol/glyceri instead of euperol?

[@Microbehunter]

Use clear nail polish. It will shrink quite a bit though, when it dries. Allow the nail polish to pre-dry or use very little of it to reduce shrinking.

[@abdulmanan370]

Wow amazing work. I'm going to stain mosquito 4th instar whole larvae for species identification and after treating them with nanoparticles kindly explain me which protocol should I follow? Reply me in comment please

[@RifaiAL]

What is the advantage of using euparal as a mounting medium? Why not use low viscosity acrylates so that you could greatly reduce the mounting time from weeks to just hours?

[@Microbehunter]

Euparal has the advantage that you can transfer the specimen from alcohol directly into it and it does not matter if there is still a bit of moisture left. Some other mounting media require the specimen to be completely free of water and they have to be placed into xylene (not healthy). Yet others are water-based. The slow drying time has the advantage in that it gives the medium time to infiltrate the specimen completely and this preserves it better. But different specimens "prefer" different media and for some resin-based mounting media might be better (this is why so many different mounting media exist).

[@bittersweet82]

​What about Ecomount as it's non toxic, is this any good?

[@WarreLattre]

Does the nail garnish work as some kind of glue so the cover glass stays in place?

[@Microbehunter]

yes. it acts both as a spacer and a glue.

[@TheRedKnight101]

How would I go about making a permanent slide of a rotifer and a ciliate?

[@Thecarverman]

@@NerdyNEET copal

[@JaySDunham]

Euparal Mounting Medium appears to no longer be available. Have you tried any of the alternatives?

[@Microbehunter]

I did find a few sources (hoping that they send it to private individuals): www.carlroth.com/en/en/Life-Science/Histology-Microscopy/Mounting-Media/Mounting-with-Euparal/Euparal/p/00000001000010c000020023_en www.agarscientific.com/mountants-euparal.html www.bioform.de/shop.php?action=tree&wg=1&pid=677&treeid=141

[@JaySDunham]

Thanks for the links! That stuff is pricey! I still want to get some, though, so I guess I'll have to bite the bullet. My German is non-existent, but that bioform web site looks interesting. Too bad there doesn't seem to be a US source for the Euparal.

[@soumyakotanoor8152]

Use Canada balsam

[@palasarinel5799]

Can Immersion Oil be used as mounting medium?

[@Microbehunter]

Yes, especially for hydrophobic specimens (eg bird feathers etc). You can also try cooking oil (cheaper).

[@computerartist7461]

What kind of alcohol do you use? You mentioned 96%, but when I'm looking at a science supply store, which kind is it? They sell Ethyl Alcohol, Isopropyl Alcohol, Methyl Alcohol, n-Butyl Alcohol etc. Thanks!

[@Microbehunter]

I use Ethyl Alcohol (ethanol). The low-cost one contains a bitter substance so that you can not drink it, and therefore there is no alcohol tax on it (Ethanol is the drinking alcohol). This is a very common one and you might even get it in a drug store or local hardware store etc. Some also call it "rubbing alcohol" but there are different compositions of this. The 99% ethanol is much more expensive and not needed. It will absorb moisture from the air until it becomes 96%. The 96% ethanol will stay like that, it is in equilibrium with the moisture in the air. Isopropyl alcohol should also work (this one is used by doctors to disinfect before giving an injection). It absorbs less moisture. For this reason it is possible to transfer the specimen from ethanol to isopropyl alcohol to completely remove water. Methyl alcohol (methanol) is VERY POISONOUS and should not be in the household. Even the fumes are dangerous. You can get blind when taking it in or even die.

[@computerartist7461]

Thank you - that is very helpful! They sell many kinds at the supply company, so this allows me to get the correct one. Also appreciate the cautions you noted.

[@computerartist7461]

Just happened to be at the drug store and they sell Isopropyl Alcohol as both 70% and 91%. Do you have a recommendation between those? For something like the insect wing, would the Isopropyl Alcohol seem to be a good choice? Thanks again.

[@Microbehunter]

Depends what you want to do with the wing. For mounting it in a mounting medium like Euparal, you need a high alcohol concentration. For storage 70% might be fine. I think that the 70% might be used for disinfection and therefore there might be water added. But maybe regular ethanol is cheaper for everyday use.

[@computerartist7461]

I had been reading the 'safe mountant' articles by Walter Dioni, and was going to try Karo syrup that he suggested for making a permanent slide. At Walmart the Isopropyl Alcohol was very inexpensive, something like 3 to 5 dollars, so I'd probably get the 91% since it has less water. Thanks!

[@TheModernVictorian]

I just found that spinner.. It is over $300 darlhurs! Nice collectable.

[@miguelelento5637]

Can I use resin?

[@Microbehunter]

The term "resin" can mean many things. Depends on which kind. I would say experiment, but make sure that you do not use toxic stuff.

[@abdulmanan370]

Which stain is it?

[@Microbehunter]

The wing was not stained, and the blue ring was made using nail polish (which is nitrocellulose, I think).

[@abdulmanan370]

@@Microbehunter thank you sir

[@elhamfarag5346]

thank you for this helpful video how can i make a permanent smear from semen sample???

[@supergamer303]

clear tape and a glass

[@par-th1]

bro i was watching this and i found a wast too

[@danielbrowniel]

You dont like killing insects? I know a guy, he's real reliable.

[@Mark-wy1ux]

How much are you paying though? I've got a great insect guy.

[@usernamemykel]

BEWARE! The "guy" is an undercover member of PITA!

[@spamcan2551]

LOL

[@spamcan2551]

@@usernamemykel that's PETA.

[@thestallion1873]

Why do you not clean your slide (before mounting) and then complain about it...you do it all the time, on all the videos...