Hello everyone,
I know that a discussion of diatom adhesives is almost like discussing religion or politics, but I have been working on the development of some "wet" diatom adhesives that I'd like to report and to get some feedback on. I can not arrange forms easily using the "dry" adhesives like Debes (gelatin/acetic acid) even when I use the tungsten needles described by Beats. The tungsten needles are wonderful but I am still unable to delicately move diatoms on the dry adhesives. Instead I have worked with moist or "wet" adhesives like Klaus Kemp used. Even though Klaus told everyone to make a tragacanth gum adhesive, he actually used a version of Meakin's (dextrine and glycerine) but added acetic acid to it which made it usable for a longer period of time. One of the problems with Klaus' (Meakin's) adhesive is that it requires you to use your thumb to wipe it on the cover (or slide which is what Klaus used). I have developed methods that don't require using your thumb!
You can read my report on the Diatom Forum site here: https://groups.io/g/diatom-forum/message/5521
Everyone should be able to read the post, but you will need to be member of the forum to see the videos that I posted in the files section.
I am very interested in finding out if these adhesives will work with Pleurax. I think they will but I am so allergic to Pleurax that I dare not try it.
Please let me know if you test them. I am very eagerly interested in other's feedback.
Thanks
Bill
I have copied my video tutorials to my Google drive so everyone should be able to see them.
Flaming a cover slip so that the adhesive will flow easily to the edges: https://drive.google.com/file/d/1knnT4ZvO7PzriLPiBX8rWN3_iBXS9gsU/view?usp=drive_link
Adding a drop of adhesive using a capillary tube onto a previously flamed cover: https://drive.google.com/file/d/1AJNlMjYS-8k3qdnoVObaUPwPXQ220OTZ/view?usp=drive_link
Let me know if you have trouble accessing these videos.
Bill
Dear Bill,
absolutely great work!
I will test it in two weeks, I have first to travel to France ;).
Thank you for sharing this here, I am sure there are a lot of happy users.
best to you
anne
Residual Oil Remover (ROR) aus den USA ist eine sehr teure Anschaffung.
Ist das von Vteroclean von OSIM nicht dasselbe, nur aus Deutschland? Viel billiger
https://www.osimoptik.de/de/Optikreiniger--VetroClean.html
Hallo Clemens,
auch Optical Wonder dürfte so ähnlich sein.
Billig sind alle nicht.
lg
anne
The heated gelatin/glycerine adhesive (cured on a hotplate at 140C for 20 minutes) can successfully be used with Pleurax as mounting media!! So the cured gelatine/glycerine adhesive can be used with BOTH toluene and alcohol-based mounting media!!
The PVA/glycerine has not been tested with Pleurax yet.
You can read the entire thread on the Diatom Forum here: https://groups.io/g/diatom-forum/topic/100319228
Bill
Hallo zusammen,
ich habe heute die Methode getestet - es funktioniert mit Pleurax.
Extrem wichtig ist, dass der Klebegrund mit den Diatomeen darauf ausreichend lange bei hoher Temperatur getrocknet wird.
Ich hatte zuerst bei ca. 130°C für 30 min getrocknet, das hat nicht ausgereicht.
Bei Trocknung bei 190-200°C für ca. 40 min wurden die Diatomeen vom Pleurax nicht mehr abgelöst.
Ob es schon bei z. B. 160°C auch funktioniert kann ich nicht sagen.
Bill Dailey, der mir im Hintergrund zur Seite stand, geht davon aus, dass die Gelatine denaturiert und dadurch nicht mehr vom Alkohol in Pleurax angegriffen wird.
Das Legen ist wesentlich einfacher da in einem Flüssigkeitsfilm leichter positioniert werden kann. Ich habe keine Reste von Kleber an den Diatomeen nach dem trocknen feststellen können.
Hello all,
I tested the method today - it works with Pleurax.
It is extremely important that the adhesive base with the diatoms on it is dried at a high temperature for a sufficiently long time.
I first dried at about 130°C for 30 min, which was not enough.
When drying at 190-200°C for about 40 min, the diatoms were no longer detached from the Pleurax.
I cannot say whether it works at 160°C, for example.
Bill Dailey, who assisted me in the background, assumes that the gelatine denatures and is therefore no longer attacked by the alcohol in Pleurax.
Laying is much easier because it can be positioned lightly in a liquid film. I have not noticed any residual glue on the diatoms after drying.
Many thanks to Bill!
anne
Dear Anne,
Thank you for testing the Debes/Beats/Downing adhesive with Pleurax since I am so allergic to it. Since Pleurax is a good, stable mounting medium and can be prepared reproducibly by a good chemist(who isn't allergic to it!), this now allows Pleurax to be used with arranged forms using the new adhesive method. It is just important to heat the gelatin enough so that it is denatured so it becomes much less soluble in alcohol. I use an alcohol flame when I cure the adhesive covers but others like you use a hotplate. Your heating temperature and time (190C for 40 minutes) is a good method for others to use. Of course it is important to measure the temperature of the hotplate so you are confident of the temperature. I use an infrared thermometer which are available for modest cost.
I will be interested and eager to hear if other people have tried this new adhesive method.
Bill
Hallo Anne , hallo Bill
Danke für das Mitteilen euerer Ergebnisse und Erfahrungen mit dem Gelatine/Glycerine Klebegrund.
Liebe Anne, ich hatte auch bei 130Grad getrocknet, auch mit negativem Ergebnis. >:( Nun erspare ich mir weitere Versuche und werde gleich nach deiner Trockenvorschrift vorgehen.
Auf meine Ergebnisse bin ich nun selbst gespannt, ob ich endlich einmal ein kleines Legepräparat mit ausreichendem Brechungsindex zustande bringe, welches auch vorzeigbar ist. :D
lg von Sigi
Hello Anne , hello Bill
Thanks for sharing your results and experiences with the gelatin/glycerine adhesive base.
Dear Anne, I had also dried at 130deg, also with negative result. >:( Now I save myself further attempts and will proceed immediately according to your drying instructions.
I am now curious about my results myself, whether I can finally produce a small laying preparation with sufficient refractive index, which is also presentable. :D
lg from Sigi
Hi Bill,
yes an Infrarot Thermoneter is one of the standard equipment to have. The setting on the hotplates is not that accurate and it is good to check it.
I checked during the curing and it was between 190°C and 197°C.
best
anne
I would also like to point out that those of you who are comfortable using the "dry" Debes gelatin adhesive should also be able to heat this to 190C for 40 min and that should allow Pleurax to be used as the mountant. I do not believe that the lack of glycerine will make any difference to the success.
If you happen to try the heating of the Debes adhesive and then use Pluerax, please post you results.
Thanks!
Bill
Hallo zusammen,
Ich habe mal eine Frage zu der Methode:
Wie sieht es mit dieser Methode aus, wenn man Diatomeen legt, aber die Arbeit nicht in einer Sitzung erledigt ist? Wie bekommt man die "Legefläche" wieder nass? Könnte mir vorstellen das das Glycerin auch irgendwann eintrocknet. Wie geht man dann weiter voran? Frisches Glycerin drauf? Nur dann besteht ja die Gefahr das die schon anhaftenden Schalen fort schwimmen, da das DG ja noch nicht auf 190 Grad erhitzt wurde.
Bei "Debes" kann man ja die Arbeit für einige Monate liegen lassen. Beim weiter arbeiten d.h. Diatomeen legen und anhauchen, ist es wieder klebrig. Und die schon geklebten Formen bleiben auch an der Stelle.
Hat da schon jemand Erfahrungen mit?
Gruss
Michael
Zitat von: Michael K. in Januar 12, 2024, 20:26:05 NACHMITTAGSWith "Debes" you can leave work behind for a few months. If you continue to work, i.e. place diatoms and breathe on them, it will be sticky again. And the shapes that have already been glued stay in place.
Does anyone already have experience with this?
Hi Michael
I once pre-coated about 40 coverslips with Debe's and stored them in a small flat cardboard box (not airtight). I wasn't making many arrangements for a while and 6 months later I still had 10 coverslips left. When I came to use one, it had lost it's "stick" (all the others had too). Breathing on it didn't moisten the Debes' anymore and diatoms didn't attach. The gelatin had dried to the point of being waterproof. Probably the same thing as heating to 190C, only done at room temperature over a much longer period.
If you plan to add more diatoms to your Diatomarium, I would suggest testing that the first form added is stuck by breathing on the mount. It may not be. Safest testing method is to pick up another diatom on your needle and use that to try and push the (hopefully) attached diatom sideways. Don't touch it with the needle though. The mounted diatom won't move if it's properly stuck. If it isn't, then best to finish the mount. Debes' works with xylene and Darax (though toluene "dissolves" any trapped bubbles faster - if you can get it).
Cheers
Beats
Hallo zusammen,
Ich habe gestern ein paar Versuche mit dem Debes Kleber und Glycerin gemacht. Also ich habe ein DG genommen und den Debes Kleber drauf gegeben, trocknen lassen. Dann einen kleinen Tropfen Glycerin drauf gegeben.
Diesen Tropfen habe ich noch mir einen winzigen Pinsel vertreilt um ein dünnen Film zu erhalten.
Ich finde es dennoch schwierig Diatomeen zu positionieren. Ich mache es ja unter dem Stereomikroskop.
Ich weis jetzt nicht wie die Sache unter dem normalem Mikroskop aussieht. Ich habe bisher kein Bild
gesehen wie es mit dem Glycerin beim legen selbst aussieht. Bei mir saugen sich die Diatomeen mit Glycerin
voll was die Erkennbarkeit erschwert.
Wie bekommt man sonst einen dünnen Glycerin Film zustande?
Nach dem Legen und troknen bei 190 Grad für ca. 40 Min und anschliessendem einschliessen (zuvor mit stark verdünndem Pleurax) mit Pleurax, zeigen das die Diatomeen an Ort und Stelle bleiben.
Hier ein Bild vom Legen mit dem Glycerin. Das Bild vom Endergebnis liefere ich nach.
LG
Michael
- Übersetzung -
Hello everyone,
Yesterday I did a few tests with Debes glue and glycerin. So I took a DG and put Debes glue on it and let it dry. Then added a small drop of glycerin.
I spread this drop over a tiny brush to get a thin film.
I still find it difficult to position diatoms. I do it under the stereo microscope.
I don't know what it looks like under a normal microscope. I don't have a picture yet
seen what it looks like with the glycerin when laying it yourself. For me, the diatoms absorb glycerin
full which makes it difficult to recognize.
How else do you get a thin glycerin film?
After laying and drying at 190 degrees for approx. 40 minutes and then enclosing (previously with highly diluted Pleurax) with Pleurax, the diatoms show that they stay in place.
Here is a picture of laying with the glycerin. I'll provide a picture of the end result later.
Greetings
Michael
Hallo zusammen,
Klaus Kemp nutzte ein evtl. ähnliches Verfahren, bis heute leider ein Geheimnis. Aber, als ich ihn besuchte zeigte er mir wie er arbeitet. Er hauchte vor jeder Diatomee das Präparat an. Dadurch war der der Klebegrund immer feucht. Ich denke auch er hat seine großen Präparate über mehrere Tage erstellt und durch das ständige Anhauchen den Klebegrund feucht gehalten.
lg
Anne
(Sorry for not translating, just sending this from the mobile)
Hallo zusammen,
Hier noch der Nachtrag bezügich der Bilder. Die Diatomeen blieben an Ort und Stelle beim eindecken.
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
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
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 (https://www.mikroskopie-forum.de/index.php?topic=48111.0)
Greetings
Michael
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
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
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
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.
Here's a better image taken on my Orthoplan using Leitz Pl Apo 25x/0.65, single image. No adhesive residue apparent.
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
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
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.