A method for rapid freeze fixation of plant cells

PROTOPLASMA ◽  
1986 ◽  
Vol 131 (2) ◽  
pp. 153-165 ◽  
Author(s):  
Susan A. Lancelle ◽  
D. A. Callaham ◽  
P. K. Hepler
Keyword(s):  
Plant Cells ◽  
Rapid Freeze Fixation ◽  
Freeze Fixation

Ultrastructural study of archaeological Vitis vinifera L. seeds using rapid-freeze fixation and substitution

2009 ◽  
Vol 41 (6) ◽  
pp. 443-447 ◽  
Author(s):  
Claudio Milanesi ◽  
Rita Vignani ◽  
Andrea Ciacci ◽  
Alessandra Nardini ◽  
Marco Valenti ◽  
...  
Keyword(s):  
Vitis Vinifera ◽  
Ultrastructural Study ◽  
Vitis Vinifera L ◽  
Rapid Freeze Fixation ◽  
Freeze Fixation

Cotton (Gossypium hirsutum) seed trichomes expand via diffuse growing mechanism

1995 ◽  
Vol 73 (5) ◽  
pp. 746-757 ◽  
Author(s):  
Suresh C. Tiwari ◽  
Thea A. Wilkins
Keyword(s):  
Gossypium Hirsutum ◽  
Cotton Seed ◽  
Morphological Changes ◽  
Plant Cells ◽  
Freeze Substitution ◽  
Diffuse Growth ◽  
Cytological Features ◽  
Freeze Fixation ◽  
Insight Into

The ultrastructure of cotton (Gossypium hirsutum) seed trichomes was investigated to obtain insight into their growth behavior during the phase of rapid cell elongation. A mold and cast method of scanning electron microscopy was used to record the morphological changes in the ovular surface during the initiation and elongation of trichomes. A rapid freeze-fixation and freeze-substitution protocol was used to study the cytological features of trichomes at 2 days after anthesis. At the cytological level, attention was primarily focused on determining whether the seed trichomes display features that are characteristic of other tip-growing plant cells, including organelle zonation, polarized deposition of cell wall, axial orientation of microtubules, and microfilament meshwork at the cell apex. Our results show that cotton seed trichomes do not share any ultrastructural characteristic with other tip-growing plant cells. Rather, they show all the characteristics of cells that undergo diffuse growth. The roles of actin microfilaments and microtubules were also investigated through an in vitro administration of cytochalasin D and colchicine. Although the disruption of actin filaments did not stop trichome growth, disruption of microtubules did prevent polarized cell expansion. Based on these results, cotton seed trichomes are not tip-growing cells but expand via diffuse growth. Key words: cytoskeleton, freeze-fixation, freeze-substitution, Gossypium hirsutum, lint fibers, tip growth, trichomes.

Rapid-freeze fixation and substitution improves tissue preservation of microspores and tapetum in Brassica napus

2000 ◽  
Vol 12 (4) ◽  
pp. 237-240 ◽  
Author(s):  
J. H. E. Ross ◽  
C. Milanesi ◽  
D. J. Murphy ◽  
Mauro Cresti
Keyword(s):  
Brassica Napus ◽  
Tissue Preservation ◽  
Rapid Freeze Fixation ◽  
Freeze Fixation

SEM study of the morphological effects of kinetin and zeatin on the growth and development of the apical meristem in wheat

1995 ◽  
Vol 53 ◽  
pp. 978-979
Author(s):  
G. M. Hutchins ◽  
J. S. Gardner
Keyword(s):  
Growth And Development ◽  
Furfuryl Alcohol ◽  
Apical Meristem ◽  
Plant Cells ◽  
Triticum Aestivum L ◽  
Morphological Development ◽  
Naturally Occurring ◽  
Chinese Spring Wheat ◽  
Sem Study ◽  
Moist Environment

Cytokinins are plant hormones that play a large and incompletely understood role in the life-cycle of plants. The goal of this study was to determine what roles cytokinins play in the morphological development of wheat. To achieve any real success in altering the development and growth of wheat, the cytokinins must be applied directly to the apical meristem, or spike of the plant. It is in this region that the plant cells are actively undergoing mitosis. Kinetin and Zeatin were the two cytokinins chosen for this experiment. Kinetin is an artificial hormone that was originally extracted from old or heated DNA. Kinetin is easily made from the reaction of adenine and furfuryl alcohol. Zeatin is a naturally occurring hormone found in corn, wheat, and many other plants.Chinese Spring Wheat (Triticum aestivum L.) was used for this experiment. Prior to planting, the seeds were germinated in a moist environment for 72 hours.

Immunocytochemistry after fast freeze fixation-freeze substitution: Advantages and limits

1990 ◽  
Vol 48 (3) ◽  
pp. 42-43
Author(s):  
Marie-Thérèse Nicolas
Keyword(s):  
Osmium Tetroxide ◽  
Freeze Substitution ◽  
Acrylic Resin ◽  
Expected Improvement ◽  
List Type ◽  
Chemical Fixation ◽  
Freeze Fixation ◽  
Liquid Chemical ◽  
Luciferase Enzyme ◽  
Lower Background

An alternative to aqueous chemical fixation consists in immobilizing physically the specimen by freezing it as fast as possible without using any cryoprotectant. This Fast Freeze Fixation (FFF) followed by Freeze Substitution (FS) avoids osmotic artefacts due to the slow penetration of liquid chemical fixative. Associated with Immuno-Gold labeling (IGS), FFF-FS allows a more precise localization of antigens.Using the bioluminescent bacteria Vibrio harveyi, a comparison of IGS with an antibody directed against its luciferase (enzyme of the luminescent reaction) has been done after liquid chemical fixation versus FFFFS. This later technique, beside an expected improvement of the ultrastructure always shows a better preservation of antigenicity and a lower background. In the case of FFF-FS technique (Figure 3):–labeling in acrylic resin (LRWhite) is 2 to 4 fold more intense than in epoxy resin (Epon),–but the ultrastructure is always better in Epon.–but the ultrastructure is always better in Epon.–The addition of fixatives in the substitution medium, results in a decrease of labeling which is more important in the case of a mixture of fixatives than with osmium tetroxide alone; with one exception: the substitution with glutaraldehyde which produces a dramatic increase in the density of the labeling but also, at the same time, a swelling of the cells of about 30%.

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