Wet Preparation

About microscopic forms of life, including Bacteria, Archea, protozoans, algae and fungi. Topics relating to viruses, viroids and prions also belong here.

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Mad_Scientist
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Wet Preparation

Post by Mad_Scientist » Sun Dec 04, 2005 6:49 am

Does anyone know what are the characteristics of a good wet preparation? Beside avoiding bubbles underneath the cover slip, what else determines a good wet preparation?

Thanks!

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Post by MrMistery » Sun Dec 04, 2005 11:26 am

Cutting it thin so you will actually see something...
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Post by Poison » Sun Dec 04, 2005 5:33 pm

And adding proper amount of water. Or it will turn into a small lake. :)
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Post by MrMistery » Sun Dec 04, 2005 8:19 pm

Actually there is no problem ifyou add more water than necessary. it won't affect the clarity of the image. The bad side of doing this is you will need to clean the table, floor etc after you are done...
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Post by Ken Ramos » Thu Dec 08, 2005 1:18 am

It would depend, I suppose, on what you are observing in the wet mount. Protozoa for example, too much water and they will swim up and down making it difficult if not impossible to maintain an accurate focus on the little buggers. To much water will also affect the image resolution at higher powers also. Preferably, just add enough water that the cover slip will not float but spread the water or other liquid equally across the surface. It will take a bit of practice to know just how much liquid to add to your preparation. :D

This Chaetogaster for example was photographed in a wet mount at 50X.
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Post by Poison » Thu Dec 08, 2005 6:34 pm

MrMistery wrote:Actually there is no problem ifyou add more water than necessary. it won't affect the clarity of the image. The bad side of doing this is you will need to clean the table, floor etc after you are done...


If you are looking at a moving thing (like a protozoa) , you can not follow it in too much water. :D
And yes, you have to clean the table. Plus the glass sometimes does not move properly because of water. It becomes like sticked.
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Post by Ken Ramos » Thu Dec 08, 2005 11:24 pm

Yes chasing after protozoa can be a testy process. However there are several things on market that can help to slow them down. One is "Proto Slow," which I have never used, wall paper paste with a high cellulose content will work also, and of course there is Lidocane, used for the numbing of a sore throat. All three of these preparations require a very small amount of the substance to have an effect on the organisms. A Tuberculin sryinge and needle work well, as does a micro pippette. :)

As for chasing them around, most higher end scopes have a mechanical stage or will accept one. These are ideal for observation after you have slowed the little critters down. The Chaetogaster was photographed using nothing at all except for the weight of the coverslip. This was an extreamly active specimen at the time of photographing, however I still was able to take high magnification images of it as well. There is also a compression device available for applying a very minute amount of pressure to the coverslip to hold things in place. :D
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Post by Poison » Fri Dec 09, 2005 7:26 am

We used a substance (I don't remember its name now) to slow down (they said it won't kill them) our protozoans in the lab. But we and the assistants were surprised after using the substance. They were all dead. :D
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Post by Ken Ramos » Fri Dec 09, 2005 11:16 am

Usually most preps that slow them down will eventually kill them but sometimes they stay alive long enough to make a sketch or two or a series of photographs. I rarely use such things anymore because the photographic set up I now use is pretty quick. So, if I catch them idle for a couple of seconds I can usually get a decent photograph. :D
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Post by MrMistery » Fri Dec 09, 2005 7:11 pm

Ok, ok... Sorry... i was thinking about plant sections...
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Post by Ken Ramos » Fri Dec 09, 2005 11:35 pm

Mr. Mistery said:
Ok, ok... Sorry... i was thinking about plant sections...


You may have me on that subject. :D I have never prepared a c.s or l.s of any of those type specimens. However I may attempt to do so in the future, if and when I acquire a microtome. They are quite expensive instruments, even used ones. :(
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Post by MrMistery » Sat Dec 10, 2005 2:22 pm

Well you don't need one. I did have one at school, but never used it since i could never open the boxes they keep them in(and neither could anyone else for that matter). Just make many sections and choose the best ones for dieing and viweing...
"As a biologist, I firmly believe that when you're dead, you're dead. Except for what you live behind in history. That's the only afterlife" - J. Craig Venter

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