Microscope, Cameras and Software[Bearbeiten]
- Light microscope Weso BM Pro1100, magnification usually 400x (Eyepiece WF 10x/Objective 40/0.65x) ->Weso
- Moticam 1000 and Moticam 5 MPixel with MoticImages V2.0/Windows 7 Professional 64 ->Moticam
- For retrieval of interesting structures on the pollen slides, I use a Pyser S7 England Finder
- CombineZP, Image Stacking Software, by Alan Hadley ->Homepage
- Picolay Image Stacking Software by Heribert Cypionka ->Homepage
- Statistical analysis with the Excel-AddIn WinStat
Pollen was prepared by staining with fuchsin solution and embedding in Kaiser glycerin gelatin. Beginning with slide 09-100 I use the prestained (0.014% fuchsin, Kaise) glycerol jelly. Melting of the gelatin by means of a USB Cup Warmer Pearl, Model 834. The coverslip was then sealed with nail polish. Degreasing of the pollen with a few drops of ether and subsequent aspiration of the liquid with filter paper.
Mix 5 ml of glycerol 85%, 10 ml of ethanol 96%, 15 ml of water and about 12 drops of a saturated solution of basic fuchsin.
Chloralcarmin-solution (CCF-staining: Pollen-Nucleus-Staining)[Bearbeiten]
Dissolve 0.2 g of carmine in 12 ml of ethanol 96% and 5 ml of hydrochloric acid 25%. Close the lid of the test tube with aluminum foil and keep it in a boiling water bath for 30 minutes. After cooling replace the evaporated ethanol and add 10 g of chloralhydrate. Filter.
Mix 20 ml of water, 20 ml of lactic acid and 20 ml of glycerol 85%. Add 20 g of phenol and 0.05 g of anilinblue (methylblue).
Glycerol gelatin (Kaiser)[Bearbeiten]
Put 7 g of gelatin (Gelatina alba Golddruck), 42 g of water, 50 g of glycerol 85% and 0.5 g of phenol in a 250 ml laboratory plastic bottle and keep it in a water bath at about 60 °C. Shake in between. If everything has dissolved, keep the bottle in the water bath until all air bubbles have accumulated on the surface. Turn the bottle slowly and fill some small 30 ml bottles with the hot solution. Allow to cool.
Glyceol gelatin (Kaiser) stained with fuchsine (0.007%)[this staining method starts with slide 09-100][Bearbeiten]
14 g of gelatin (Gelatina alba Golddruck), 84 g of water, 100 g of glycerol 85%, 0.5 g of phenol and 14 mg fuchsine (C.I. 42510, z.B. Merck 1.15937)
Glycerol gelatin (Kisser)[Bearbeiten]
10 g of gelatin, 35 g of water, 30 ml of glycerol 85% and 0.5 g of phenol.
Dissolve 16 g of chloralhydrate in 10 g of water and add 5 ml of glycerol 85%.
Farbstoff zur Pollenfärbung. Der Farbstoff ist direkt in der Glyceringelatine (nach Kaiser, Fuchsin 0.007%) gelöst.