Experiments with botanic stains

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MicroBob
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Location: Northern Germany

Experiments with botanic stains

#1 Post by MicroBob » Tue Dec 08, 2020 3:59 pm

Hi together,
in former times there was a stain called "Kernschwarz" of unknown formulation. It stained unlignified cell walls very specifically. Today it is not available any more but it is thought to be based on gallic acid and an iron salt, like iron gall ink.
This made me think how iron gall ink might behave as part of a botanic stain. The one I bought has a strong added blue component (where iron gall ink would normaly be black after oxidisation on the paper).
This here is so far the most promising result:
5 drops water
1 drop ink
3 drops SFC solution (Safranine-diamon fuchsine-chryosidine 1%, 1% 0,5%)

Applied for 8 hours, washed with water, dried with isopropanol and mounted in Euparal.

Slide was fresh, please excuse the uneven sharpness!

Bob
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5-1-3 Wasser-Salix-SFC DSC_1024.jpg
5-1-3 Wasser-Salix-SFC DSC_1024.jpg (330.26 KiB) Viewed 1236 times

Wes
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Joined: Sat Mar 09, 2019 12:58 pm

Re: Experiments with botanic stains

#2 Post by Wes » Tue Dec 08, 2020 4:27 pm

Impressive results Bob! Is this a hand section, looks great. Also which component gives the yellow color?
Zeiss Photomicroscope III BF/DF/Pol/Ph/DIC/(epi)FL/Jamin-Lebedeff
Youtube channel

MicroBob
Posts: 2544
Joined: Sun Dec 25, 2016 9:11 am
Location: Northern Germany

Re: Experiments with botanic stains

#3 Post by MicroBob » Tue Dec 08, 2020 4:29 pm

Hi Wes,
this was cut embedded in a slit carrot with a cylinder microtome with my 3D-printed blade holder. The leaf sectioned very well.
The SFC-mix geves the yellow, the ink the black and blue tones.

Bob

Wes
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Joined: Sat Mar 09, 2019 12:58 pm

Re: Experiments with botanic stains

#4 Post by Wes » Tue Dec 08, 2020 4:40 pm

Do you have a link where your microtome and sectioning technique are described?
Zeiss Photomicroscope III BF/DF/Pol/Ph/DIC/(epi)FL/Jamin-Lebedeff
Youtube channel

MicroBob
Posts: 2544
Joined: Sun Dec 25, 2016 9:11 am
Location: Northern Germany

Re: Experiments with botanic stains

#5 Post by MicroBob » Tue Dec 08, 2020 5:18 pm

I have showed my hand /cylinder microtome and the blade holder I have developed here. I actually have a hand microtome for sale.

viewtopic.php?f=16&t=7065&p=62601&hilit ... der#p62601

deBult
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Joined: Tue Jul 16, 2019 3:20 pm

Re: Experiments with botanic stains

#6 Post by deBult » Tue Dec 08, 2020 8:04 pm

I’m using Bob’s knife holder and counter: strongly recommended (and well worth the extra world wide shipping cost).

deBult
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Joined: Tue Jul 16, 2019 3:20 pm

Re: Experiments with botanic stains

#7 Post by deBult » Tue Dec 08, 2020 8:11 pm

The differentiation in tissue types looks quite good to me Bob.

Iike the overall balance in colours as well.

MicroBob
Posts: 2544
Joined: Sun Dec 25, 2016 9:11 am
Location: Northern Germany

Re: Experiments with botanic stains

#8 Post by MicroBob » Tue Dec 08, 2020 8:33 pm

In the german forum the Wacker W3A variations are very popular, actually have been varied, developed and improved by a couple of members. These are very very good stains but as well as Etzold FCA they are quite colourful, over time may be a bit overly colourful. When Rolf-Dieter-Müller, one of the staining experts, showed a Kernschwarz-Safranin stain in the black-orange range I was hoping that this would become available as a recipe based on todays available substances. This month we will have a virtual group meeting on botanic staining and hand sections so this gave me the idea to contribute a bit to the development (if ony as a bad example).

Im' more the Pareto principle type trying to get 80% with 20% effort and be happy with it. So this simple simultaneous stain would be quite nice if it prooves durable and usable.

Here another image where I removed the blue tint with disodiumdisulfit. But I start to like the blue version better...

Bob
Attachments
5-1-3 Wasser-Salix-SFC Dinatriumdisulfit DSC_1016 1024.jpg
5-1-3 Wasser-Salix-SFC Dinatriumdisulfit DSC_1016 1024.jpg (316.55 KiB) Viewed 1198 times

Chris Dee
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Joined: Mon Aug 12, 2019 1:02 pm

Re: Experiments with botanic stains

#9 Post by Chris Dee » Wed Dec 09, 2020 10:50 am

Nice experimental stain work Bob. Thanks for sharing.

Hobbyst46
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Joined: Mon Aug 21, 2017 9:02 pm

Re: Experiments with botanic stains

#10 Post by Hobbyst46 » Wed Dec 09, 2020 12:40 pm

Hi Bob,
I like the first one, the bluish section. Very nice !
Zeiss Standard GFL+Canon EOS-M10, Olympus VMZ stereo

MicroBob
Posts: 2544
Joined: Sun Dec 25, 2016 9:11 am
Location: Northern Germany

Re: Experiments with botanic stains

#11 Post by MicroBob » Wed Dec 09, 2020 2:11 pm

Hi Doron,
I was looking for a black-orange stain, but start to prefer the blueish one too.
What I found is that bleaching the section with Clorix changes the apperance of the staining a lot.

Bob

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