Saharan spontaneous plants are very lignified this lignification makes the histological section very difficult, I tried to find the best methods of allocation of cuts for these plants.
Hi Noureddine,
To get nice sections on lignified tissues you have several strategies depending on what information you research.
For a structural information, you can fix your samples into Carnoy's fixative, then embed your samples into a hard resin such as methacrylate (LR White or Technovit 7100) or epoxy (Spurr, Epon or Araldite) and cut blocks with a glass, saphire or diamond knife.
You will have to cut very thin sections less than1µm thick.
If you are looking for immunodetection, a formaldehyde fixative followed by emmbedding in LR White is fine but immuno is tricky then.
Does this answer ?
Cheers
Oliv
yes thank you. because in my work I used solid paraffin, i can't get nice section
Hello,
Perhaps you could try thin grinding of your specimens.
It is a technique which is used in bone histology and also geology. But I believe it also would give good results with lignified tissues.
Specimens are embedded into plastic (for instance Technovit 7200 VLC), cut in half with a diamond coated saw, glued to a slide and then ground to a thickness of 50 to 100 µm. You can polish and stain them after that.
A pdf which describes how it is done can be downloaded from the following link:
http://www.ganam.co.kr/sboard/download.php?id=data&no=1&gubun=1
You cannot do immunochemistry with these specimens but you obtain optimal structural integrity.
The machines and technique are rather expensive, but for starters you could perhaps try to grind them by hand on commercial grinding and polishing paper.
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