CRYO-PLANING FOR CRYO-SCANNING ELECTRON MICROSCOPY (SEM) -A METHOD FOR OBSERVATION OF XYLEM CONTENTS IN ROSA STEMS

A sample preparation technique for low temperature scanning electron microscopy, called Cryo-Planing for Cryo-Scanning Electron Microscopy (Cryo-SEM) is described.
Successful cryo-planing required a stable attachment of the specimen to a holder.
Clamping with a screw mechanism and using indium as space filler sufficed for 3 cryo-planing methods studied:
1. cryo-milling
2. microtomy in a cold room
3. cryo-ultramicrotomy
From the 3 methods, the latter one was preferred.
A small modification of this cryo-planing method using cryo-ultramicrotomy was then applied for a series of Cryo-SEM observations.
The results showed that the cryo-ultramicrotomy technique enables observations of flat sample surfaces of frozen, fully hydrated plant specimen such as stems of ornamental flowers -roses and chrysanthemum-, but also for Cryo-SEM observations of hazel stems, other plant materials like rapeseed pistils and even dermatological research samples.
The cryo-ultramicrotomy technique can be applied successfully:
a. to improve observations from cryo-SEM specimen,
b. to enhance results from x-ray microanalysis,
c. to enhance results from (digital) image analysis.
The planing method can be applied on fibrous, hard, and delicate materials as well.

IV International Symposium on Rose Research and Cultivation

J. Nijsse, H.M.C. Put

https://doi.org/10.17660/ActaHortic.2007.751.62