ISHS Acta Horticulturae 751: IV International Symposium on Rose Research and Cultivation
CRYO-PLANING FOR CRYO-SCANNING ELECTRON MICROSCOPY (SEM) -A METHOD FOR OBSERVATION OF XYLEM CONTENTS IN ROSA STEMS |
| Authors: | J. Nijsse, H.M.C. Put |
| Abstract:
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. | |
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