Rose stem

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bernhardinho
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Joined: Sun Aug 13, 2006 6:28 am
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Rose stem

Post by bernhardinho »

Hi folks,

I did practice my thin section cutting routine yesterday. So here is a rose stem at 100x . The dye is astrablue/safranin/chrysoidin. The third image is under crossed polars and compensator red I, Coolpix 990.

Image

Image

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Greetings

Bernhard

Bruce Williams
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Post by Bruce Williams »

Bernhard,

Section cutting and dying is still some time in the furure for me. These are wonderfully detailed images that will have me chasing off to the web to learn the basics of plant structure.

Did you prepare the sections as permanent mounts?

Bruce

Ken Ramos
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Location: lat=35.4005&lon=-81.9841

Post by Ken Ramos »

Bernhard :!: Now we are getting down to some serious stuff here. I would have to say that you are the first that I can recall, of actually having cut and stained a specimen for post. From what I can see you are really good at it IMO. I have not advanced to that degree of expertise but have often given it thought however. What sort of microtome did you use in cutting the c.s.? :D

gpmatthews
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Post by gpmatthews »

Beautiful - I agree with Ken, must be one of the first, if not THE first with your own cut and stained sections posted in the forum. Now that's what you call cut flowers!
Graham

Though we lean upon the same balustrade, the colours of the mountain are different.

bernhardinho
Posts: 563
Joined: Sun Aug 13, 2006 6:28 am
Location: Germany
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Post by bernhardinho »

Hi folks

thanks for your overwhelming response. I would like to emphasize that I am a total beginner in this buisness and I encourage everybody to give it a try, There are a lot of traps but very soon you get the hang of it and will be able to achieve quite reasonable results.

The technique:

The stem has been fixed in 70% Ethanol. I then put the stem in between two halfs of a carrot which then is inserted in a hand cylinder (or bench) microtome. The actual cutting has been performed using a special razor blade fastened in a holder. The thin sections then went into a bleeching bath using Eau de Javelle, the staining I mentioned can be purchased as a standard dye called Etzold's mixture. After the staining, which takes a couple of minutes, the sections need to get differenciated in 70% Ethanol which draws out excess stain. before mounting the specimens need to be dehydrated in 100% Isopropanol (hope that's the english word as well). All these procedures I do on a slide. When the 100% alcool has nearly evaporated I put on a drop of euparal (mounting resin) and finally the cover slip. Let it dry out and that's it.

Just try it, they look even nicer in real life than my poor art of photography can show.

Bernhard


P.S. By the way, Bruce, look here for a start : http://en.wikipedia.org/wiki/Plant_stem

Walter Piorkowski
Posts: 691
Joined: Mon Aug 14, 2006 6:42 pm
Location: South Beloit, Ill

Post by Walter Piorkowski »

My congratulations to you on your images. I have long wanted to make thin sections of plant material but fell into the trap of thinking that I had to follow the whole dehydration and wax impregnation process as well as the final sectioning using a microtome. I began to test that whole process after aquiring the materials but stopped due to the amount of time required. I have made pleasing "thick" sections however.

Your work here has given me a push to try this work again. How thick was your section when it was put on the slide? How long was your initial 70% ethanol fix? I will try your process.

Walt

Walter Piorkowski
Posts: 691
Joined: Mon Aug 14, 2006 6:42 pm
Location: South Beloit, Ill

Post by Walter Piorkowski »

My congratulations to you on your images. I have long wanted to make thin sections of plant material but fell into the trap of thinking that I had to follow the whole dehydration and wax impregnation process as well as the final sectioning using a microtome. I began to test that whole process after aquiring the materials but stopped due to the amount of time required. I have made pleasing "thick" sections however.

Your work here has given me a push to try this work again. How thick was your section when it was put on the slide? How long was your initial 70% ethanol fix? I will try your process.

Walt

bernhardinho
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Joined: Sun Aug 13, 2006 6:28 am
Location: Germany
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Post by bernhardinho »

Hi Walter,

well if you take something sturdy like a reasonably sized plant stem, there is no need to embed it. I collected the stem a couple of weeks ago and kept it in the alcool, thus being able to practice every now and then. How thin the section actually was I cannot tell, to be honest, certainly not thin enough (probably around 70µm) . but this process of bleaching (sorry about my wrong spelling above) does a good job removing the cell contents and you can see the nice grid. ( By the way, you can use one of those bleaching agents used for cleaning the bathroom ; Klorix being the name of a brand over here in Germany.)

Best wishes

Bernhard

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