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Posted: Sun Nov 21, 2021 9:28 pm
by Microscopy_is_fun
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Re: Hello from Austria

Posted: Sun Nov 21, 2021 9:54 pm
by Dubious
Welcome! Beautiful images. The cross section of mistletoe resembles a mosaic work of art.

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Posted: Sun Nov 21, 2021 10:03 pm
by Microscopy_is_fun
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Re: Hello from Austria

Posted: Sun Nov 21, 2021 10:21 pm
by charlie g
Welcome to a terrific community/online forum ( thank you, Oliver Kim). Did you process the two botanical stained sections you share with us in this post? Wonderful sections. Please share with us the workup process and stain schedule for these two plant sections.

Please use this forums search feature to enjoy the botanical microtechnique of member: "Mr.Sonchas" .

I have purchased wonderful stained plant slides from Mr.Sonchas..please yourself 'take a peek' at his low cost/high quality botanical slides.

welcome, welcome...err: 'bruderschaft microscopy'!

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Posted: Sun Nov 21, 2021 10:50 pm
by Microscopy_is_fun
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Re: Hello from Austria

Posted: Sun Nov 21, 2021 11:45 pm
by charlie g
Wow, both your sections seem a lot thinner than: '30-40 microns' the 'bare spots' in the mistletoe section..clear of 30-40 microns column of tissue..yet with crisp defined cell walls...and the crisp nuclei in the pine needle section seem too clear of 'above/ beneath tissues' to be 30-40 micron sections'..great achievement, thanks for sharing your process.

welcome, charlie g

Re: Hello from Austria

Posted: Mon Nov 22, 2021 9:05 am
by Hobbyst46
Welcome, what nice micro images ! I like the auto fluorescence image of the needle !

And the microtome is impressive - most of it is made from standard industrial profiles I see. I believe many would find interest in the construction details.

Re: Hello from Austria

Posted: Mon Nov 22, 2021 12:49 pm
by Javier
Welcome!

Those images are quite impressive. I'm looking forward to seeing more of your work.

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Posted: Mon Nov 22, 2021 7:19 pm
by Microscopy_is_fun
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Re: Hello from Austria

Posted: Mon Nov 22, 2021 7:28 pm
by Wes
Welcome! Your photos are really good and that microtome looks pretty amazing too. Looking forward to more of your posts :)

Re: Hello from Austria

Posted: Mon Nov 22, 2021 11:04 pm
by mete
Great images. May I ask the procedure for PEG 1500 embedding ? It is preferred over higher molecular weights because it is a weak tissue or for other reasons ?

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Posted: Tue Nov 23, 2021 9:24 pm
by Microscopy_is_fun
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Re: Hello from Austria

Posted: Wed Nov 24, 2021 7:22 am
by mete
Microscopy_is_fun wrote:
Tue Nov 23, 2021 9:24 pm
@mete: I took the original recipe for PEG from the internet, unfortunately I forgot the source. I slightly modified it, and use the following procedure now:

(1) fixing the samples in AFE
(2) rinsing in aqua dest
(3) around 6h in 20wt%PEG 1500/80wt% aqua dest
(4) around 4h in 50%PEG/50%aqua dest.
(5) around 6h in PEG 1500 @55°C
(6) casting a PEG block with the embedded sample for slicing
(7) after slicing dissolution of PEG in 20wt% PEG / 80wt% aqua dest
(8) staining and mounting as desired

It is important to replace the water in the sample gently with PEG: water diffuses faster than PEG, and if the exchange process happens to fast (e.g. by skipping the 50/50-stage or putting the slices directly into aqua dest.), the sample gets easily destroyed.

Overall a fairly easy process, involving no problematic solvents or other nasty chemicals.
Thanks, I think I will try PEG, I have a few questions but I will create a new post.