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Glycerol Permeabilization of Fluorescently-Labeled Phalloidin Improves Actin Visualization in Vibratome-Sectioned Roots

Published online by Cambridge University Press:  02 July 2020

E.B. Blancaflor  and
K.H. Hasenstein
E.B. Blancaflor
Affiliation:
Plant Biology Division, The Samuel Roberts Noble Foundation Inc., 2510 Sam Noble Parkway, Ardmore, OK, 73401
K.H. Hasenstein
Affiliation:
Department of Biology, University of Louisiana, P.O. Box 42451, Lafayette, LA, 70504
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Extract

The actin cytoskeleton is involved in various cellular functions including cell shape determination, organelle transport, and cell polarity establishment. In order to study the function and dynamics of actin in plant cells, methods to image its orientation have to be optimized. Although, green fluorescent proteins (GFP) from the jellyfish Aequoria victoria have been used for visualizing actin filaments in plant cells, the technique is still in its infancy and has some limitations. Actin in plant cells is difficult to preserve and hence visualize mainly because of the problems associated with specimen fixation and processing. This is particularly true for bulky specimens such as plant organs because of the additional requirement for embedding and sectioning after the tissues are fixed and dehydrated. In order to overcome these difficulties, we have modified a method previously used for onion epidermal cells and suspension cells, and applied this to fresh Vibratome sectioned root tissues.

Type
Biological Specimen Preparation/Cytochemistry/ Immunolabeling/Immunocytochemistry
Information
Microscopy and Microanalysis , Volume 6 , Issue S2: Proceedings: Microscopy & Microanalysis 2000, Microscopy Society of America 58th Annual Meeting, Microbeam Analysis Society 34th Annual Meeting, Microscopical Society of Canada/Societe de Microscopie de Canada 27th Annual Meeting, Philadelphia, Pennsylvania August 13-17, 2000 , August 2000 , pp. 470 - 471
Copyright
Copyright © Microscopy Society of America

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References

References:

1.Kost, B. et al., Plant J. 16 (1998) 393.CrossRefGoogle Scholar
2.Olyslaegers, G. and Verbelen, J.P., J. Micros. 192 (1998) 73.CrossRefGoogle Scholar
3.Blancaflor, E.B. and Hasenstein, K.H., Plant Physiol. 113 (1997) 1447.CrossRefGoogle Scholar
4. This work was supported by the Samuel Roberts Noble Foundation, Inc. (E.B.B.) and the National Aeronautics and Space Administration (NASA grant NAG 10-0190 to K.H.H.).Google Scholar