We will need some chemicals and some equipment:
Isopropyl pure
Ethanol 70%
Ethanol 50%
Ethanol 30%
Aqua dest.
Etzold Staining Solution (Astra Blue, Fuchsine, Chrysoidine)
Fixation Solution AFE (90% Ethanol, 5% Formalin, 5% Glacial Acid)
Euparal
Pipette: 4x
Watch Glas
The general procedure will be to leave the cuts in the watch glas and remove the chemicals with a pipette. Use 1 pipette for AFE, 1 for Ethanol, 1 for Aqua dest and stains, 1 for Isopropyl. It is very important that the Isopropyl is as dry as possible. Water in the Isopropyl will remove the Chrysoidine. It is a good idea to shake the watch glas a little bit after a new chemical is applied.
Fill the watch glas (or another small container with a similar form) with AFE. Now cut fresh material either by hand or a small microtome. Each section that is cut will be put into the watch glas. After the last cut is safely in the watch glas, simply follow this procedure:
15 min AFE, then remove with pipette 1
5 min Ethanol 70%, then remove with pipette 2
3 min Ethanol 50%, then remove with pipette 2
3 min Ethanol 30%, then remove with pipette 2
3 min Aqua dest, change 3 times with pipette 3
10 min Etzold Staining Solution, apply heat (50° C), then remove with pipette 3. Use 10 drops of the staining solution so the sections are covered with the staining solution.
Use Aqua dest. to rinse of excess staining solutions, remove with pipette 3
10 sec Isopropyl pure, remove with pipette 4
2 min Isopropyl pure, remove with pipette 4
5 min Isopropyl pure, remove with pipette 4
Isopropyl pure
Now use a Stereo Microscope or something similar to find the best samples and put them with a fine brush onto a freshly cleaned slide. Put a drop of Euparal on it and a cover glas. Apply some pressure (careful!).
Now bake at 40° C for 48 hours. Voila, we are done. If you did everything correct the result will look like this:
English Yew, Taxus baccata
Give it a try and show your results. It is nowhere as difficult as it sounds (even I can do it) and a lot of fun once you mastered it.
Questions are always welcome and if you need help obtaining the Etzold Staining Solution, please drop me a Message and I will help.
Good luck!
