"Wet" diatom adhesives - looking for testers and feedback

[Michael K.]

Hallo zusammen,

Hier noch der Nachtrag bezügich der Bilder. Die Diatomeen blieben an Ort und Stelle beim eindecken.

[bill2penn]

Hello Michael,
Very good results so far with Pleurax.
To get a very thin film of glycerine you will need to dilute it strongly with solvents such as acetic acid and alcohols(isopropanol and isobutanol). I will provide my current recipe for the glycerine/Debes below. I think it would be easiest for you to apply both Debes and glycerine at the same time as I do rather than try to add a glycerine cover to the already placed Debes coating.

I make "stock" solutions of
0,3g gelatin in 1 mL water and then add 25 mL acetic acid
2 g of glycerine in 10 mL acetic acid
12 mL isopropanol and 6 mL isobutanol

To make the glycerin/Debes adhesive I mix
2 parts gelatin stock solution
1 part glycerine stock solution
2 parts pure acetic acid
then slowly with good mixing add 2 parts of the alcohols.

I add a very small drop of this to a previously cleaned and flamed coverslip and let dry for at least 30 minutes, but I have let it go overnight.

There should be a very, very thin film of liquid adhesive that should not completely cover the diatom forms but you should be able to see the liquid on the tips. If you have a lot of liquid on the cover after you let it dry, try to add smaller drops or add additional acetic acid and alcohols in equal amounts to make it more dilute.
If required, you can breathe on the cover to remoisten it.

Let us know how you make out!
Best regards,
Bill

[bill2penn]

Hello again Michael,

You should try to use the regular Debes to arrange a few diatoms (if you are comfortable moving forms on dry surface) and then heat this to 190C. Then try with Pleurax. The glycerin may not be needed.

Bill

[Michael K.]

Hello Bill,

Thanks for your answer. Unfortunately, I didn't know that you could dilute glycerin. I'll try the recipe out.

Here is the link to the "dry" laying experiment with Pleurax.
https://www.mikroskopie-forum.de/index.php?topic=48111.0


Greetings
Michael

[Michael K.]

Hallo zusammen,


Es wäre auch für andere hilfreich, wenn man auch Bilder vom präparieren selber sehen könnte.
Ich habe jetzt mal das Original Rezept von Bill angewendet. Er schreibt ja das man Flüssigkeit oder den feuchten Klebegrund in den Ecken / Spitzen an der Diatomeen sehen sollte.
Ich habe hier mein ein Bild, wie so etwas aussehen kann. Es scheint etwas zuviel zu sein, aber bei meinem letzten Präparat wo die Diatomeen komplett mit reinem Glycerin "abgesoffen" sind und später mit Pleurax eingedeckt wurden, waren die im Mikroskopbild ordentlich zu sehen. Ich wusste da nicht wie man Glycerin "richtig" verdünnt.
Mich und vermutlich auch anderen, würde es interessieren wie die Diatomeen im nassen Medium beim legen auszusehen haben.
Ich werde das Präparat einfach mal ferig legen und dann mal sehen wie es später mit Plx aussieht.

Gruss
Michael


Hello everyone,


It would also be helpful for others if you could see pictures of the preparation yourself.
I've now used Bill's original recipe. He writes that you should see liquid or the moist adhesive base in the corners/tips of the diatom.
Here I have a picture of what something like this might look like. It seems to be a bit too much, but in my last specimen where the diatoms were completely "soaked" with pure glycerol and were later covered with Pleurax, they were clearly visible in the microscope image. I didn't know how to dilute glycerin "correctly".
I, and probably others too, would be interested to see what the diatoms look like when laid in the wet medium.
I'll just finish the preparation and then see what it looks like with Plx later.

Greeting
Michael

[bill2penn]

Hi Michael,
I think this is good but you will have to heat and see if any adhesive can be seen on the form. You should "push" the forms on the adhesive so they get "wet" adhesive on them. If you simply place a form on the adhesive and it becomes completely wet, then that is probably too much liquid. But small, thin forms are sometimes covered with liquid.

To get a thiner film of liquid you can either use a smaller drop, or perhaps easier is to dilute your adhesive with more acetic acid/alcohol mixture.

You can see how Klaus Kemp did this with his wet adhesive in this video https://www.youtube.com/watch?v=zrK3ncElKm0  between 2:10 and 3:00

Bill

[bill2penn]

Here is an image of several forms in the "wet" adhesive along with my tungsten needle. You can just see a little liquid next to the tip of the needle and the small piece of dirt. It's difficult for me to take a good image showing the liquid. But I will try again later.
Bill

[bill2penn]

And here is the finished slide after heating to set the adhesive and mounting in Zrax. This is a single image using the same objective and single image using a 20X objectove. I should note that the centric form had very obvious liquid in the center but this was removed by the heating. And there doesn't seem to be any gelatin adhesive that can be seen on any of the forms.

[bill2penn]

Here's a better image taken on my Orthoplan using Leitz Pl Apo 25x/0.65, single image. No adhesive residue apparent.

[Michael K.]

Hallo zusammen,


Ersteinmal Danke für die Bisherigen Antworten !
Ich habe heute Nachmittag mal ein kleines Präparat gelegt, um zu testen wie es sich mit wet Mount arbeiten lässt.
Die Diatomeen habe ich aufgesetzt und verschieben können, wo es nötig war. Angedrückt habe ich nicht alle,
vor allem die kleinen nicht. Aber auch hier blieben die Schalen nach Zugabe von Pleurax an Position.
Bilder von den einzelnen Schritten habe ich unten angehängt. Diese sind per Handy am Okular gemacht worden. 

Vielen Dank nochmal an Prof. Dailey für die Bemühungen und Bereitstellung des Rezeptes.


Gruss
Michael 


Hello everyone,


First of all, thanks for the answers so far!
This afternoon I made a small preparation to test how it works with wet mount.
I set up the diatoms and were able to move them where necessary. I didn't press all of them,
especially not the little ones. But here too, the shells remained in position after adding Pleurax.
I have attached pictures of the individual steps below. These were taken using a cell phone on the eyepiece.

Many thanks again to Prof. Dailey for his efforts and providing the recipe.


Greeting
Michael

[bill2penn]

Hello Michael,
Beautiful work!!

In my previous note I meant to say that you should "slide" or "move sideways" the forms on the slide so that they passed through the wet adhesive as Klaus does in the video. I did not mean to "press down" on them which may break them.

I am delighted that this method seems to work well with Pleurax mountant.

Best regards to all,
Bill

[Michael K.]

Hello,

Yes, I think it's a good method. But I think it's important that you heat it well.
It might be good if others could stop their attempts too.


Greeting
Michael


Edit:
Google translation translated "push" as "press down" in message #20 (in German). Hence the slight misunderstanding.