A few botanic cross sections

[daruosha]

Hi everybody,

cross section of pathos leaf stem (or petiole).

4X Achro Na 0.10- Condenser Aperture 0.10 - 10 Images stacked in PS
Safranin and Methylene Blue staining


cross section of a succulent stem

4x Plan Achro NA 0.10 - Condenser Aperture 0.10
Safranin and Alcian Blue staining

[BramHuntingNematodes]

Neat! What are we looking at here exactly?

[MicroBob]

Hi Daruosh,
nice even sections! The first one could have been a Sansevieria, right?

Bob

[daruosha]

Thanks Bob and BramHuntingNematodes.

I'm sorry that I didn't explain anything at all. I did all the cutting, staining, mounting and photography in one single session (nearly 5 hours) and lost my energy at the very last minute of posting the pictures.

The first image is a cross section of pathos leaf stem (or petiole).

The second image is cross section of a succulent stem (not sure about the exact specie, i'll post pictures of the actual plants for better identification).

[BramHuntingNematodes]

Beautiful sections. I would be interested in a rundown of your microtechniques.

[MicroBob]

It appears as if your stains have mixed and formed a new colour instead of attatching to different structures. Have you tried the stains alone and have you tried to differntiate them by removing part of the staining so lightly stained structures are destained?

Bob

[daruosha]

It appears as if your stains have mixed and formed a new colour instead of attatching to different structures. Have you tried the stains alone and have you tried to differntiate them by removing part of the staining so lightly stained structures are destained?

Bob

I found the exact same problem during my photography session. Before applying each stain I washed the sample in alcohol bath, I don't know what is the problem, but certainly I did something wrong and I would like to re-run the whole thing once again in a more control fashion. The quarantine thingy started to melt my brain down...

[daruosha]

Beautiful sections. I would be interested in a rundown of your microtechniques.

Thanks BramHuntingNematodes for your kind comments. I'm in the process of writing a rather comprehensive document about my sample preparation and a survey of all the techniques I've learned so far. Will start a serial topic very soon on that subject.

[KD Arvidsson]

Great pictures Daruosh! Thanks //KD

[shirleey22]

which microscope was used?

[DonSchaeffer]

I can never get them neat!

[deBult]

If you can read the German language: the microgroup Bonn has a treasure full of cutting and staining technique info plus a description on what plant tissue structures you are actually observing.

http://www.mikroskopie-bonn.de/index.php

Best, deBult

[daruosha]

Great pictures Daruosh! Thanks //KD

Thank you KD.

which microscope was used?

It's a cheap Chinese no name microscope. I've made a few modification. but it's nothing fancy or special at all.

[daruosha]

I can never get them neat!

DonSchaeffer , yes you definitely can do it. It's nothing hard or difficult, it's time consuming and all you need is passion. As I mentioned earlier, I'm writing a guide about sample preparation and will start a separate topic.

If you can read the German language: the microgroup Bonn has a treasure full of cutting and staining technique info plus a description on what plant tissue structures you are actually observing.

http://www.mikroskopie-bonn.de/index.php

Best, deBult

That website is truly a treasure. MicroBob recommended that website earlier and I'm still reading it. There's so much to learn.

[shirleey22]

Great pictures Daruosh! Thanks //KD

Thank you KD.

which microscope was used?

It's a cheap Chinese no name microscope. I've made a few modification. but it's nothing fancy or special at all.

it looks amazing honestly! <3

[daruosha]

it looks amazing honestly! <3

Thanks for your nice comments shirleey22. I assume you have similar swift microscope. Try some specimens and show us your pictures.

[shirleey22]

it looks amazing honestly! <3

Thanks for your nice comments shirleey22. I assume you have similar swift microscope. Try some specimens and show us your pictures.

i had a Swift SW350B but had to return it. I’m expecting my SW380B and i sure will show you some. Thank you!

[MicroBob]

Safranin was part of the old Etzold FSA stain. It was replaced with Chryosidine because this is staining in a bit more differentiated way. But Etzold wouldn't have used it if it were not suitable.
Alcian blue is part of the Wacker W3A stain so it is sure that it is good for plant sections, Wacker new his stains.

Generally some stains are applied until the staining intensitiy is right while others are applied intensively and then revoved as necessary. But I'm no expert here. The FSA and W3A protocols would be a good startig point, but you would just use the one stain out of three you have.

Here is an interesting comparison of staining processes: http://www.mikroskopie-bonn.de/biblioth ... k/303.html
And here the new RDM5 stain which makes use of two of your stains: http://www.mikroskopie-bonn.de/biblioth ... k/300.html
I just got it yesterday but haven't tried it yet. Apparently it gives a nicely differentiated staining in agreeable colours. Acridine red is not longer available and Rhodamine is only a so-so replacement.
This here is a very beautiful stain: https://www.mikroskopie-forum.de/index. ... ic=33452.0 Unfortunately "Kernschwarz" has always been a mystic component and the production process has never been published - and it is no longer available.

Bob

[daruosha]

Excellent tips Bob.

Tomorrow I'll do another run and try my best to document my process. Hopefully the result will be much better. I was rather successfully in the past when it comes to staining botanic cross sections. My biggest problem is that I don't follow my exact protocol (I like to take shortcuts) and last time my laziness bit me. I'll try to stick to correct staining periods/timings.