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Visualisation of microtubules and actin filaments in fixed BY-2 suspension cells using an optimised whole mount immunolabelling protocol.
- Source :
-
Plant cell reports [Plant Cell Rep] 2006 Aug; Vol. 25 (8), pp. 758-66. Date of Electronic Publication: 2006 Mar 10. - Publication Year :
- 2006
-
Abstract
- Excellent visualisation of microtubules and actin filaments was obtained in fixed tobacco BY-2 suspension cells after optimising a protocol for whole mount immunolabelling. The procedure is based on modification of fixation, cell wall digestion, dimethyl sulfoxide (DMSO) treatment, post fixation, and blocking. The most critical aspects of successful preservation and visualization of cytoskeletal elements appeared to be: a two-step fixation with paraformaldehyde and glutaraldehyde before enzymatic cell wall digestion and a post fixation with aldehydes thereafter. The method allows the improved visualization of the organisation of the microtubular and actin filament arrays during the successive stages of cell division and at interphase. Although we present the application of our protocols for cytoskeleton labelling, the excellent results show the potential of using this method for the analysis of various proteins and molecules in plant cells.
Details
- Language :
- English
- ISSN :
- 0721-7714
- Volume :
- 25
- Issue :
- 8
- Database :
- MEDLINE
- Journal :
- Plant cell reports
- Publication Type :
- Academic Journal
- Accession number :
- 16528566
- Full Text :
- https://doi.org/10.1007/s00299-005-0089-y