Bamboo (images added)

Images made through a microscope. All subject types.

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Ecki
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Bamboo (images added)

Post by Ecki »

Two weeks ago I was supposed to hold a workshop on cutting and staining fresh botanical material. As I had been occupied with protists so far this year, I decided the evening before the workshop to practice a bit and cut something from our garden. My choice was the Bamboo as this is a Monocotyledon and has vascular bundles everywhere. When I approached the Bamboo I was a bit afraid of the very hard sprouts so I settled for a fresh sprout.

I was shocked when I saw the result. It looked as if there was one color missing. It was to late to brew fresh paints and I did not sleep very well, wondering how the workshop would work if my paints fail.

For the workshop I demonstrated cutting and staining with a fern and everything went well. All colors were present as they were supposed to. While driving home it dawned on me that my fresh sprout simply did not contain lignin! Otherwise it could not be used as food because it is well known that humans don't digest wood well ;) The absence of lignin caused my staining technique to fail miserably.

As a matter of fact, most Monocotyledons produce very little lignin as they get their stability through the vascular bundles - vertical pipes providing the structural support. Because of the absence of lignin they can bend with the wind. Bamboo develops lots of lignin at a later stage and is quite sturdy.

Enough said, this is a stained section of the Garden Bamboo Fargesia sp.. This was a well lignified sprout. Enjoy!

Image
5x polarized light

Image
10x polarized light

Image
10x bright field

The next image is pretty interesting. It is well known that some large organic molecules show fluorescence as way to get rid of abundant energy:

http://en.wikipedia.org/wiki/Autofluorescence

This is a fresh cut with water under the coverslip. No chemicals applied! This is reflected light with a 100 Watt halogen lamp and the Zeiss Filter 9. The filter has an excitation wavelength of 450-490 nm (blue light) and lets emitted light with a wavelength greater 515 nm pass.

Image
20x, chlorophyll is red, lignin is green

Best regards,
Ecki
Last edited by Ecki on Tue Jun 19, 2012 11:33 am, edited 2 times in total.

rjlittlefield
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Post by rjlittlefield »

I enjoy the images very much, and the story of the lignin-free bamboo is great! I'm sorry to hear about the lost sleep, but all's well that ends well. :D

--Rik

Pau
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Post by Pau »

Ecki,
wonderful technique and pictures, and an interesting story behind of course
Pau

RogelioMoreno
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Post by RogelioMoreno »

Ecki,

Very well done and great story. :D


Rogelio

SteveGreen1953
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Post by SteveGreen1953 »

Most interesting.

abpho
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Post by abpho »

Very cool.

Cactusdave
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Post by Cactusdave »

Excellent work. What compensator(s) are you using in combination with your polariser to achieve these particularly striking colours in the polarisation shots?
Leitz Ortholux 1, Zeiss standard, Nikon Diaphot inverted, Canon photographic gear

Mitch640
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Post by Mitch640 »

Your stain slice pictures are always so amazing. Beautiful.

discomorphella
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Post by discomorphella »

Great autofluorescence image. I am always glad to see informative and beautiful fluorescence images obtained without the use of special fluorochromes.
David

Ecki
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Post by Ecki »

Good that you like those sections. More will be coming as the summer progresses :D

This is an image of the sections that I prepared before the workshop:

Image
10x

The colors look like the paints did not work. My expectation was to have colors like this:

Image
10x

This is a section of a well lignified sprout. The same staining procedure has been applied to both sections. Now you can image my confusion ...

The last image is in polarized light of the section without lignin:

Image
2,5 Pol, details enhanced with software

@Dave, this was polarized darkfield. I suppressed some of the red (which is dominant on those lignified sprouts) by amplifying the blue channel with software.

Regards
Ecki

Chris S.
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Post by Chris S. »

Arresting images, and very interesting analysis/story, Ecki. Thanks!

--Chris

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