caa a22 4500 475817451 CHVBK 20180406123812.0 cr unu---uuuuu 170329e20000101xx s 000 0 eng 10.1007/s004970050007 doi (NATIONALLICENCE)springer-10.1007/s004970050007 Rapid-freeze fixation and substitution improves tissue preservation of microspores and tapetum in Brassica napus [Elektronische Daten] [J. H. E. Ross, C. Milanesi, D. J. Murphy, Mauro Cresti] Abstract : The method of rapid freeze-fixation and substitution was used with Brassica napus floral bud material in order to improve the preservation of microspore and tapetal organelle structure. When observed using transmission electron microscopy, the appearance of the freeze-substituted material differs in a number of ways from the chemically-fixed material previously studied, in particular for the lipid-rich elaioplasts and tapetosomes in the tapetal cells. The tapetosomes have a very electron-dense, opaque appearance when visualized after rapid fixation. In addition, we were able to observe other cytoplasmic details such as pockets in the endoplasmic reticulum and cytoskeletal structures such as microfilaments. Extracellular material was also well-preserved; for example, the fibrous material in the baculae of the developing microspore exine was also visible. Finally, in the freeze-fixed sections specific structures such as elaioplasts could be labelled by antibodies, which indicates that this method preserved protein epitopes that were destroyed by chemical fixation. Springer-Verlag Berlin Heidelberg, 2000 Key words Freeze-substitution nationallicence Transmission electron microscopy nationallicence Brassica napus nationallicence Tapetum nationallicence Microspore nationallicence Immunogold nationallicence Ross J. H. E. Department of Brassica and Oilseeds Research, John Innes Centre, Colney Lane, Norwich, NR4 7UH, UK, GB aut Milanesi C. Dipartimento di Scienze Ambientali, Università degli Studi di Siena,Via P.A. Mattioli, 4, I-53100, Siena, Italy, IT aut Murphy D. J. Department of Brassica and Oilseeds Research, John Innes Centre, Colney Lane, Norwich, NR4 7UH, UK, GB aut Cresti Mauro Dipartimento di Scienze Ambientali, Università degli Studi di Siena,Via P.A. Mattioli, 4, I-53100, Siena, Italy, IT aut https://doi.org/10.1007/s004970050007 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s004970050007 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Ross J. H. E. Department of Brassica and Oilseeds Research, John Innes Centre, Colney Lane, Norwich, NR4 7UH, UK, GB aut NATIONALLICENCE 700 1- Milanesi C. Dipartimento di Scienze Ambientali, Università degli Studi di Siena,Via P.A. Mattioli, 4, I-53100, Siena, Italy, IT aut NATIONALLICENCE 700 1- Murphy D. J. Department of Brassica and Oilseeds Research, John Innes Centre, Colney Lane, Norwich, NR4 7UH, UK, GB aut NATIONALLICENCE 700 1- Cresti Mauro Dipartimento di Scienze Ambientali, Università degli Studi di Siena,Via P.A. Mattioli, 4, I-53100, Siena, Italy, IT aut Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer