I’m struggling to have a closer look at Rotifers.

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Jonnyvine
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I’m struggling to have a closer look at Rotifers.

#1 Post by Jonnyvine » Wed Mar 07, 2018 9:17 am

Hello fellow microscopists.

I have been recently looking at Rotifers that I’ve cultured, but there is one problem. They seem to move around a lot and I struggle to get a good view on 400x magnification to have a closer look at them.

Is there a way slow them down or freeze them in a way? :|

Lilly Begonia
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Re: I’m struggling to have a closer look at Rotifers.

#2 Post by Lilly Begonia » Wed Mar 07, 2018 9:33 am

Jonnyvine wrote:Hello fellow microscopists.

I have been recently looking at Rotifers that I’ve cultured, but there is one problem. They seem to move around a lot and I struggle to get a good view on 400x magnification to have a closer look at them.

Is there a way slow them down or freeze them in a way? :|
Well nearly all of the rotifers I've come across have been dormant, and only a very few were moving at all, but i get your problem. I see lots of micro-organisms that are moving around so fast I can't get any decent shot of them. It's pretty frustrating having to chase them all over the slide getting nothing but bad pictures. I frankly do not know the answer to it.

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75RR
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Re: I’m struggling to have a closer look at Rotifers.

#3 Post by 75RR » Wed Mar 07, 2018 9:50 am

charlie g's slow'm down recipe: 50/50 (half specimen fluid + half 1.5% concentration methyl cellulose)

This is the stuff: https://www.homesciencetools.com/produc ... ose-30-ml/
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Hobbyst46
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Re: I’m struggling to have a closer look at Rotifers.

#4 Post by Hobbyst46 » Wed Mar 07, 2018 1:13 pm

Thanks 75RR, good to know about a commercial methyl cellulose solution for this purpose. Last time I used glycerol but MC is definitely better.
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Micro-Bob
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Re: I’m struggling to have a closer look at Rotifers.

#5 Post by Micro-Bob » Wed Mar 07, 2018 4:09 pm

Your water layer might be too thick. You can remove some water by drawing it out with a piece of cloth.

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Jonnyvine
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Re: I’m struggling to have a closer look at Rotifers.

#6 Post by Jonnyvine » Wed Mar 07, 2018 4:19 pm

It's not that because I can see them swimming and moving all around the slide. When they are not swimming their tail moves like caterpillars.

There was a rare moment where they stood still for 10secs and open their mouth.

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Re: I’m struggling to have a closer look at Rotifers.

#7 Post by einman » Wed Mar 07, 2018 8:10 pm

You can "slow them down" using methylcellulose or anesthetize them using various agents such as magnesium chloride, Lidocaine, Neosynephrine among others.

Older publications mention Chloral Hydrate and cocaine although those chemicals are prohibitive in our day and age for obvious reasons.
Last edited by einman on Wed Mar 07, 2018 10:05 pm, edited 1 time in total.

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Re: I’m struggling to have a closer look at Rotifers.

#8 Post by photomicro » Wed Mar 07, 2018 8:34 pm

Some medicines in the form of a spray for numbing the throat contain lidocaine and can slow creatures down by just adding a little to the water (start low, you can always add more).

However, a compressorium of some sort is often best. Look this up to see what it is. There are various types, and you can make your own.

Mike

Hobbyst46
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Re: I’m struggling to have a closer look at Rotifers.

#9 Post by Hobbyst46 » Wed Mar 07, 2018 10:50 pm

photomicro wrote: However, a compressorium of some sort is often best. Look this up to see what it is. There are various types, and you can make your own.
Mike
Thanks for the interesting information. Google led me to David Walker's article in Micscape about a Victorian-era version of this instrument with its medieval-flavor name...
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einman
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Re: I’m struggling to have a closer look at Rotifers.

#10 Post by einman » Thu Mar 08, 2018 12:58 am

Good luck finding one! I have tried for a year now.

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Re: I’m struggling to have a closer look at Rotifers.

#11 Post by charlie g » Thu Mar 08, 2018 3:37 am

Hi, jonneyvine ( BTW, I grow concord grapes in finger lakes/US )....absurd to reach for delicate pharmacy drugs...or silly and bulky 'gadgets'...placed on your microscopes stage...like...err...'compresoriums'...when low cost...yes cheap...viscosity tool : methylcellulose is so handy a 'viscosity tool' for your higher magnification observations of rotifers, water mites, water fleas, nematodes, water bears ( all these and more termed: meiofauna)...and for the usually smaller: protists ( protozoa...and yes, the larger and quite mobile bacteria).

Thanks, 75RR, for the excellent link to a firm offering low cost ( cheap and wonderful viscosity tool!) methylcellulose.

Please, pretty please, jonnyvine, use 50/50 sample fluid and methylcellulose as 75RR kindly advised you to try with your microscopy?! Enjoy observing the excretory 'flame cells' gentle lambent flicker in your rotifer subjects by this wetmount slide simple setup.

As has been suggested in this great thread you posted...keep the fluid thickness under your wetmount slides coverslip...a thin volume...think of it as a 'sweet spot' of water colum thicknes...simply feed more water to coverslip border when evaporation dries down the waters under your slides cover slip. Please online, or with a text...look at sketches of rotifer flame cells...and yourself experience this profound microscopy encounter!

Burried in this great forum are my 'shout outs' to use of : viscosity tool methylcellulose for live protist and meiofauna 'normal behaviours..yet slowed down'. Charlie guevara

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Jonnyvine
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Re: I’m struggling to have a closer look at Rotifers.

#12 Post by Jonnyvine » Thu Mar 08, 2018 9:07 am

Is there a cheaper method? Something I can find around the home?

photomicro
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Re: I’m struggling to have a closer look at Rotifers.

#13 Post by photomicro » Thu Mar 08, 2018 9:23 am

Jonnyvine wrote:Is there a cheaper method? Something I can find around the home?
Yes, mix up a tiny amount of wall-paper paste to a thin consistency. If it works, and you want to keep it, you might need to add something to stop mould growing, but it is cheap enough to make as and when you need it.

A compressorium is a generic term, doesn't have to be fancy, or expensive, and you can easily make your own. It might not be as good as some of the purpose made ones, but...

Another technique...if you put three or four small blobs of vaseline on the slide, within the cover-slip area, you can press the cover down a little, and if you have a small water drop, this may also restrict the rotifer.

There are some good old publications by Galliford, Hollowday and others, about practical issues with rotifers.

Note also, that bdelloid rotifers are quite different from some of the planktonic types, and need different techniques to view easily.

Have fun.

Mike

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Jonnyvine
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Re: I’m struggling to have a closer look at Rotifers.

#14 Post by Jonnyvine » Thu Mar 08, 2018 9:38 am

Thanks for the great advice photomicro! :D

Will try out wallpaper paste later this week.

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Re: I’m struggling to have a closer look at Rotifers.

#15 Post by SATCOM » Wed Mar 14, 2018 6:48 pm

Johnnyvine,

Philodina is the most common rotifer to be found locally and is easy to locate. I have found that the best way to view protozoa is to have lots of them to look at, I concentrate them. Collect large of amounts of pond vegetation and rinse it in bucket with the water. Collect several batches and rince and drain each. Keep a couple more bunches in the bucket. Back home divide up bottom water (full of detritus) into a few jars, add water and vegetation to each. As the jars settle things concentrate. When removing a small amount of the lower level water/detritus, slowly use pipette and allow this to settle also before adding drop to slide. I’ve even put 50ml tubes and centrifuge for a 10-15 seconds. I like to use well slides for the increased volume of water. These double rotors guys can be found happily attached to detritus sucking up food. Easy to see the little mastic parts crunching food intact.

Local pound and slow moving pond.also provides water mites ( love the blue ones with red spots), daphnia, clam shrimps, cyclops, scuds, paramecium, sun animalcules (ameba with spikes seen first by van Leeuwenhoek), tested (amber donut empty tests) and naked amebas, vorticella is an attached rotifer, desmids, water bears, and many many other interesting critters, plants (sprirogrira and be scanned to see sex and the tested amoeba difugulia eating it), and unknowns.

Sorry for getting off track, but bottom concentrated water will get you rotifers.

SATCOM

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coominya
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Re: I’m struggling to have a closer look at Rotifers.

#16 Post by coominya » Sun Mar 18, 2018 10:06 am

Jonnyvine wrote:It's not that because I can see them swimming and moving all around the slide. When they are not swimming their tail moves like caterpillars.

There was a rare moment where they stood still for 10secs and open their mouth.
I had that experience the other day. I gave up observing and went out shopping, then visiting, and hours and hours later I came back and turned on the microscope lamp and they were all very immobile :)

There is probably a sweet spot, where they are slow but not dead.

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Jonnyvine
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Re: I’m struggling to have a closer look at Rotifers.

#17 Post by Jonnyvine » Mon Mar 19, 2018 8:59 am

The problem is that if they are dead, you can't see them opening their mouth when they feed.

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Jonnyvine
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Re: I’m struggling to have a closer look at Rotifers.

#18 Post by Jonnyvine » Thu Mar 22, 2018 9:01 am

photomicro gave a nice idea about using wallpaper paste. Will that stuff wash off the slides?

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Re: I’m struggling to have a closer look at Rotifers.

#19 Post by Peter » Thu Mar 22, 2018 4:36 pm

Jonnyvine wrote:photomicro gave a nice idea about using wallpaper paste. Will that stuff wash off the slides?
Yes it will wash off the slides.

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