wax necessary for microtome?

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njitgrad
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wax necessary for microtome?

#1 Post by njitgrad » Thu Feb 18, 2016 2:39 pm

I just purchased a microtome for my 10 year old son and a lot of the tutorial videos I have been watching on YT show the use of melted wax to obtain thin cross-sections of plant tissue. Is this the recommended approach? Any recommendations on the equipment required to make and apply melted wax on demand?

This is the microtome we have:
http://www.hometrainingtools.com/microtome

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rnabholz
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Re: wax necessary for microtome?

#2 Post by rnabholz » Thu Feb 18, 2016 3:01 pm

For a hand microtome, you have some options.

My favorite "carrier" is carrot. See this thread for some pictures of the process.

viewtopic.php?f=6&t=2290&hilit=Carnation

You can sandwich the sample like shown, or I have also used a drill bit to make a channel when your material to be sectioned is long, like a stem. Make the channel as close to the size of the material as possible, a tight fit makes much better sections.

Let me know if you have questions.

Rod

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Re: wax necessary for microtome?

#3 Post by njitgrad » Thu Feb 18, 2016 3:18 pm

rnabholz wrote:For a hand microtome, you have some options.

My favorite "carrier" is carrot. See this thread for some pictures of the process.

viewtopic.php?f=6&t=2290&hilit=Carnation

You can sandwich the sample like shown, or I have also used a drill bit to make a channel when your material to be sectioned is long, like a stem. Make the channel as close to the size of the material as possible, a tight fit makes much better sections.

Let me know if you have questions.

Rod
Thanks for the thread. That really helped me out a lot. Since my microtome has a clamp as well this can easily be repeated.
BTW, do you rinse the microtome and blade with water after using them?

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Re: wax necessary for microtome?

#4 Post by rnabholz » Thu Feb 18, 2016 3:45 pm

njitgrad wrote:
rnabholz wrote:For a hand microtome, you have some options.

My favorite "carrier" is carrot. See this thread for some pictures of the process.

viewtopic.php?f=6&t=2290&hilit=Carnation

You can sandwich the sample like shown, or I have also used a drill bit to make a channel when your material to be sectioned is long, like a stem. Make the channel as close to the size of the material as possible, a tight fit makes much better sections.

Let me know if you have questions.

Rod
Thanks for the thread. That really helped me out a lot. Since my microtome has a clamp as well this can easily be repeated.
BTW, do you rinse the microtome and blade with water after using them?
Yes, always clean everything up when you are done. Depending on the material you are cutting, you may find that you need to clean the blade and microtome table at some point in the middle of the process. Cutting pine needles and pine scales was one of those times for me. Pine sap does not make the blade slide easily. ;^)

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Re: wax necessary for microtome?

#5 Post by mrsonchus » Thu Feb 18, 2016 6:47 pm

Rod said it all - you won't need wax for this microtome, indeed it isn't your best option either - carrot probably is.
Good luck, looking forward to seeing your sections. :)
John B

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Re: wax necessary for microtome?

#6 Post by njitgrad » Thu Feb 18, 2016 7:22 pm

mrsonchus wrote:Rod said it all - you won't need wax for this microtome, indeed it isn't your best option either - carrot probably is.
Good luck, looking forward to seeing your sections. :)
Thanks, you've all been a great help.

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Re: wax necessary for microtome?

#7 Post by JimT » Thu Feb 18, 2016 10:14 pm

Good luck, looking forward to seeing your sections.
Me too.

I usually just stuff more "Things" in the hole since I don't have a side clamp. Now that I have seen your microtome I am going to do some shopping.

JimT

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Re: wax necessary for microtome?

#8 Post by rnabholz » Thu Feb 18, 2016 10:51 pm

I have the same microtome and meant to mention a tip earlier that would have saved me some time and aggravation.

In my early attempts, I was having trouble with inconsistent thickness in my sections. I ultimately discovered it was because the mechanism of this microtome will allow the entire internal clamp and the enclosed sample to move upward during your cut unless you do one very simple thing.

Once you advance the sample to the desired cut thickness using the bottom dial, grasp the microtome, and simply place your thumb on the top side of the clamping screw and apply light pressure downward toward the bottom of the microtome.

That simple maneuver will insure that the sample cannot move upward during your cut. If you don't do that, the knife attack angle or even your other fingers can move the sample upward and you will find your sections uneven and thick.

I don't consider this a defect at all, just part of the protocol that should be followed when using this microtome. I like mine very much.

Good luck

Rod
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Re: wax necessary for microtome?

#9 Post by p3aul » Fri Feb 19, 2016 3:28 am

This thread is very interesting! I don't have a hand microtome yet but I want one like the one like you have. I have gone beyond onion and want to see the cells of other plants. Do you mind if I borrow your thread for a little while and ask a couple of questions?

If I just put a plant stem in length-wise I think all I would get would be a cross-section. To see plant cells I guess I would have to find some way to mount the stem side ways in the microtome. Is that correct? Is their a better procedure? What about the cells that make up a leaf? Oops that's three questions! Sorry! ;)

Thanks,
Paul
Paul Microscope: Amscope T400b Camera: Amscope MU300
Telescope: Orion xt6 classic Dob, Zhumell z10 classic Dob

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rnabholz
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Re: wax necessary for microtome?

#10 Post by rnabholz » Fri Feb 19, 2016 2:15 pm

p3aul wrote:This thread is very interesting! I don't have a hand microtome yet but I want one like the one like you have. I have gone beyond onion and want to see the cells of other plants. Do you mind if I borrow your thread for a little while and ask a couple of questions?

If I just put a plant stem in length-wise I think all I would get would be a cross-section. To see plant cells I guess I would have to find some way to mount the stem side ways in the microtome. Is that correct? Is their a better procedure? What about the cells that make up a leaf? Oops that's three questions! Sorry! ;)

Thanks,
Paul
Sectioning a leaf in any direction is very simple, you simply sandwich the dimension you are interested in between the carrot halves and cut.

Here is a thread showing cross and longitudinal cuts on a philodendron leaf:

viewtopic.php?f=6&t=636&hilit=Philodendron

A longitudinal cut on a stem does present a challenge. If I were to want to try it, I would probably use a channel in a carrot, and fit the carrot very closely to the diameter of the microtome's throat. Then make cuts down the length until I reached the sample.

Hopefully the side of the throat of the microtome would hold the sample in place. The cuts would work only until you reached the center of the sample, after that the stem would no longer be held by the carrot.

This would likely only work with a fairly stiff stem and a sharp knife.

Perhaps others will offer a better approach.

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Re: wax necessary for microtome?

#11 Post by njitgrad » Fri Feb 19, 2016 8:07 pm

rnabholz wrote:Once you advance the sample to the desired cut thickness using the bottom dial, grasp the microtome, and simply place your thumb on the top side of the clamping screw and apply light pressure downward toward the bottom of the microtome.
Thanks for the tip!

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Re: wax necessary for microtome?

#12 Post by njitgrad » Fri Feb 19, 2016 8:09 pm

rnabholz wrote:Yes, always clean everything up when you are done. Depending on the material you are cutting, you may find that you need to clean the blade and microtome table at some point in the middle of the process.
Just rinse under water and prop the microtome on a towel at an angle to dry?

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Re: wax necessary for microtome?

#13 Post by JimT » Fri Feb 19, 2016 8:34 pm

Perhaps others will offer a better approach.
I have only done it once and what I did was to cut the stem length so it would fit in the microtome. The hole in my microtome is small so the stem fit without the need of a carrot. Based on the picture you provided you will want to use a carrot to wedge the stem in place. Lots of trial and error until you get a cut thin enough but worth the effort.

Good luck.

JimT

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Re: wax necessary for microtome?

#14 Post by rnabholz » Fri Feb 19, 2016 8:48 pm

njitgrad wrote:
rnabholz wrote:Yes, always clean everything up when you are done. Depending on the material you are cutting, you may find that you need to clean the blade and microtome table at some point in the middle of the process.
Just rinse under water and prop the microtome on a towel at an angle to dry?
Yes, hot water and dish soap if needed. Then I just set mine, table down on a towel and give it overnight to dry. Water could accumulate in the bottom of the tube and perhaps cause some issues otherwise.

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Re: wax necessary for microtome?

#15 Post by rnabholz » Fri Feb 19, 2016 8:52 pm

JimT wrote:
Perhaps others will offer a better approach.
I have only done it once and what I did was to cut the stem length so it would fit in the microtome. The hole in my microtome is small so the stem fit without the need of a carrot. Based on the picture you provided you will want to use a carrot to wedge the stem in place. Lots of trial and error until you get a cut thin enough but worth the effort.

Good luck.

JimT
Thanks for jumping in Jim.

It occurred to me since my earlier post that it would be easier to simply not make the channel go all the way through the carrot, which would then stop the sample from moving as the knife pressed against it during the cut.

I will have to give that a go to test the theory.

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