biologia plantarum

Biologia plantarum 15:50-56, 1973 | DOI: 10.1007/BF02922513

On the media improving freeze-sectioning of plant material

K. BeneŠ¹

¹ Institute of Experimental Botany, Czechoslovak Academy of Science, Praha 6-Vokovice, Czechoslovakia

Using a semiconductor freezing microtome, there is no difficulty in getting an ample supply of quite suitable sections of Ca formol fixed root tips ofVicia faba, applying Holt's sirup or plain 0.88 M sucrose as media for keeping the material, getting the sections and treating them after sectioning, but the protoplasts shrink and often fall out from the cells. No improvement in this respect was revealed using - at different concentrations - sucrose and arabic gum alone. The changes in the proportions of the ingredients of the sirup were also ineffective. A series of experiments were therefore performed with plain sucrose, but none of the factors tested was efficient enough to improve the results as compared with the standard procedure, the gradual transfer being the only exception. Since this makes the procedure more laborious and time consuming, certain other media were tested, 0.88 M ethanol and 0.88 M dimethylsulf-oxide yielding the best results. Attempts are being made to understand some points of the present experience in connection with the results of other authors and on the basis of general theory respectively.

Received: March 14, 1972; Published: January 1, 1973  Show citation