Ciliate in Methylcellulose

Here you can post pictures and videos to show others.
Post Reply
Message
Author
Chris Dee
Posts: 191
Joined: Mon Aug 12, 2019 1:02 pm

Ciliate in Methylcellulose

#1 Post by Chris Dee » Wed Oct 30, 2019 3:29 pm

Odd reaction to Methylcellulose of a ciliate (1% solution, 50/50 mix with sample water). Other ciliates on the slide showed normal behaviour, just slowed down as expected. Found others on a second slide also exhibiting this pulsing motion of the oral groove/gullet. Is this normal?


mnmyco
Posts: 144
Joined: Tue Aug 28, 2018 11:03 pm

Re: Ciliate in Methylcellulose

#2 Post by mnmyco » Wed Oct 30, 2019 5:26 pm

I wonder if happened to get a very small particle methylcellulose that did not dissolve and is trying to clear it.

Chris Dee
Posts: 191
Joined: Mon Aug 12, 2019 1:02 pm

Re: Ciliate in Methylcellulose

#3 Post by Chris Dee » Wed Oct 30, 2019 6:01 pm

The sample was well mixed but possibly. Perhaps some ciliates find it an irritant?

mintakax
Posts: 270
Joined: Mon Jul 29, 2019 12:06 am
Location: Boulder CO, USA

Re: Ciliate in Methylcellulose

#4 Post by mintakax » Wed Oct 30, 2019 7:23 pm

Chris-- Was this a commercial preparation ? The reason I ask is I have purchased MC from two different companies and both have killed most of the protists within minutes. This video shows what happened, second half of vid shows rupture in cell membrane:

https://vimeo.com/368305002

I contacted the second company.. they watched the video, ran some tests of their own and told me that they have identified a problem with the substance.

Chris Dee
Posts: 191
Joined: Mon Aug 12, 2019 1:02 pm

Re: Ciliate in Methylcellulose

#5 Post by Chris Dee » Wed Oct 30, 2019 9:07 pm

I didn't notice any increase in mortality over the base sample water mintakax. There were only a few exhibiting this behaviour over 4 examined slides, all the others were going about their business happily at reduced speed. Purchased from eBay, it was advertised as 'Refined Methylcellulose' from an established trader with good feedback. I expect if not buying analytical grade from a big laboratory supplier, there's always a risk of contamination.
Edit: Was purchased as a powder, mixed by me with boiled/cold rainwater.

mintakax
Posts: 270
Joined: Mon Jul 29, 2019 12:06 am
Location: Boulder CO, USA

Re: Ciliate in Methylcellulose

#6 Post by mintakax » Wed Oct 30, 2019 10:53 pm

Chris Dee wrote:
Wed Oct 30, 2019 9:07 pm
I didn't notice any increase in mortality over the base sample water mintakax. There were only a few exhibiting this behaviour over 4 examined slides, all the others were going about their business happily at reduced speed. Purchased from eBay, it was advertised as 'Refined Methylcellulose' from an established trader with good feedback. I expect if not buying analytical grade from a big laboratory supplier, there's always a risk of contamination.
Edit: Was purchased as a powder, mixed by me with boiled/cold rainwater.
Thanks. I think the powder is the way to go. The stuff I had was already mixed and had salicylic acid as a preservative and thats what I think did the damage.

Chris Dee
Posts: 191
Joined: Mon Aug 12, 2019 1:02 pm

Re: Ciliate in Methylcellulose

#7 Post by Chris Dee » Wed Oct 30, 2019 11:50 pm

In my random testing of household stuff for relaxant properties, salicylic acid also didn't fair well. Conversely if commercial outlets are adding preservatives it does make me wonder about the storage life of diy solutions.

mnmyco
Posts: 144
Joined: Tue Aug 28, 2018 11:03 pm

Re: Ciliate in Methylcellulose

#8 Post by mnmyco » Thu Oct 31, 2019 1:56 am

I have asked this company for a sample, but I also had it shipped to my work, at a university. Their samples are 500 g.

https://www.cpkelco.com/products/cellulose-gum-cmc/

I have CMC and gellan gum from them. Works well.

MNMynco

Hobbyst46
Posts: 3034
Joined: Mon Aug 21, 2017 9:02 pm

Re: Ciliate in Methylcellulose

#9 Post by Hobbyst46 » Thu Oct 31, 2019 12:24 pm

Methyl cellulose (MC) and similar chemicals (CMC) are known and well established. However, there is another (commercial) possibility: A relatively benign powder named Polyox WSR301. It has been studied in an article from 1977 by Dr. DM Spoon, and was shown to excel in slowing the motion of protozoa; furthermore, it was shown to be non-toxic and better than MC for this purpose.
WSR301 has a molecular weight of millions, so it does not change the osmotic pressure. Ir seems to be available through Amazon.
Zeiss Standard GFL+Canon EOS-M10, Olympus VMZ stereo

mintakax
Posts: 270
Joined: Mon Jul 29, 2019 12:06 am
Location: Boulder CO, USA

Re: Ciliate in Methylcellulose

#10 Post by mintakax » Fri Nov 01, 2019 10:09 pm

Hobbyst46 wrote:
Thu Oct 31, 2019 12:24 pm
Methyl cellulose (MC) and similar chemicals (CMC) are known and well established. However, there is another (commercial) possibility: A relatively benign powder named Polyox WSR301. It has been studied in an article from 1977 by Dr. DM Spoon, and was shown to excel in slowing the motion of protozoa; furthermore, it was shown to be non-toxic and better than MC for this purpose.
WSR301 has a molecular weight of millions, so it does not change the osmotic pressure. Ir seems to be available through Amazon.
I did a search for Polyox on Amazon (USA)-- the search results are .... well..... a bit different :oops: :shock:

Hobbyst46
Posts: 3034
Joined: Mon Aug 21, 2017 9:02 pm

Re: Ciliate in Methylcellulose

#11 Post by Hobbyst46 » Fri Nov 01, 2019 10:32 pm

mintakax wrote:
Fri Nov 01, 2019 10:09 pm
Hobbyst46 wrote:
Thu Oct 31, 2019 12:24 pm
Methyl cellulose (MC) and similar chemicals (CMC) are known and well established. However, there is another (commercial) possibility: A relatively benign powder named Polyox WSR301. It has been studied in an article from 1977 by Dr. DM Spoon, and was shown to excel in slowing the motion of protozoa; furthermore, it was shown to be non-toxic and better than MC for this purpose.
WSR301 has a molecular weight of millions, so it does not change the osmotic pressure. Ir seems to be available through Amazon.
I did a search for Polyox on Amazon (USA)-- the search results are .... well..... a bit different :oops: :shock:
Sorry, I was wrong about Amazon. So I did some literature search, where did various researchers get their WSR301 (Polyox) from in recent years. The WSR301 and similar products are all Dow Chemical products; there is a supplier - Colorcon - apparently based in both the USA (in West Point - no further details) and the UK. Possibly they sell or even donate sample quantities.
Zeiss Standard GFL+Canon EOS-M10, Olympus VMZ stereo

charlie g
Posts: 1477
Joined: Mon Oct 20, 2014 7:54 pm

Re: Ciliate in Methylcellulose

#12 Post by charlie g » Sun Nov 03, 2019 6:47 pm

Thanks Chris for this excellent MC thread, posts quite thoughtful and specific for my needs. Vance Tartar in his 1961 text on Stentor species opines..different protozoa have different sensitivities to MC immersion, but Vance uses his MC solutions no more than 2 months, than makes fresh batch, Vance claims to have left Stentors for two days in MC with protozoa returning to normal when washed free of the MC.

I am cheapo and purchase low cost 1.5%MC , use it with a dedicated needle probe as applicator, alcohol wipe down applicator immediately after it's use..return probe to it's plastic tube sheath, treat that MC vial with same care to lessen contamination. Err...cheapo me, I use the vial of MC for years! If I felt the need, I would purchase MC more often.

Relis B.Brown, 1944 advises: 'place a ring of MC 5% on your microscope slide, place the organism sample droplet inside this ring...then place coverslip.'. Dr.Brown notes you achieve a gradient of MC viscosity effects by this method...near total immobile...to near free motions of protozoa.

It was Douglas A.Marsland ( published 11/5/1943!!) who advised 10% MC to 'slow down paramecia for oil-immersion lens studies'.

Dr. Vance Tartar in 1960 attached fine cloth mesh fibers to a microscope slide, attached the fine mesh with melted paraffin wax. Dr.Tartar plopped a glob of MC onto this fiber matt..this his 'surgical table' for intricate cutting/ grafting/ alterarations of large Stentor in regeneration, and morphology regeneration studies. His subject individually plunged into the MC glob..and the delicate surgeries performed on the slide prep. Use of MC, it's prep, culture of protozoa all describe in the classic 1961 text: "The Biology of Stentor".

thanks for this thread forum folks, Charlie guevara

Chris Dee
Posts: 191
Joined: Mon Aug 12, 2019 1:02 pm

Re: Ciliate in Methylcellulose

#13 Post by Chris Dee » Sun Nov 03, 2019 9:22 pm

Thanks for the info' Charlie, very useful. I like the Relis B.Brown recommendation and will try this ring method out. If I can get it to work well enough it would avoid having to apply MC to the whole sample, my only concern is how much it will elevate the cover slip. I recently bought a batch of cheap 50ml needle applicator bottles from ebay, the type with a retained silicon stopper. I like them, they minimise air and surface contact and have decanted my stains, mountant, MC, etc, to them now.

Thanks to everyone for the feedback and suggestions.

charlie g
Posts: 1477
Joined: Mon Oct 20, 2014 7:54 pm

Re: Ciliate in Methylcellulose

#14 Post by charlie g » Mon Nov 04, 2019 12:54 am

Hi Chris , I totally agree with our tight working distance / high magnification objectives, there is a sweet spot amount of sample fluid+ MC...to keep the slide/sample film/coverslip sandwich at very thin thickness. Often in wetmount slides, the fluid 'moats up' all around the cover slip border zone...so right off you have a threatening zone to avoid with high mag objectives!

I suggest large rectangular coverslips permitting scan and observations within 80% of the slide ..far from coverslip borders...just a thought. Charlie guevara

Post Reply