Effects of Different Fixatives on Freeze-Fractured Samples of Plant Cells for SEM Observations

Three-dimensional structures of meristematic cells of Allium cepa were studied using freeze-fracture method under the scanning electron microscope. Two fixation procedures were used. The cells were often fractured between eytoplasm and nucleus when the materials were fixed in 1% OsO4 alone before freeze fracture, whereas the nuclei, were frequently fractured if the materials were fixed first in Carnoy's, solution (ethanol: acetic acid=3:l) and then in 1% OsO4 before freeze fracture. The former fixation procedure is suitable for the study of the interior structures of cytoplasm such as cytoskeleton fibres, mitochondria, endoplasmic reticulum and their three-dimensional topography. The latter fixation method is suitable for the study of interior structures of nucleus such as chromatin, nucleoli, nuclear matrix filaments and their 3-dimensional architectures, especially the 3-dimensional structures of chromatin in fibrillar centre of the nucleolus.     

L'ultrastructure de l'adénohypophyse du crapaud Bufo bufo L.

Résumé Les types cellulaires de l'adénohypophyse du Crapaud (Bufo bufo L.) ont été identifiés et étudiés avec deux fixateurs différents. La fixation au tétroxyde d'osmium permet l'identification de cinq types de cellules, alors que celle à la glutaraldéhyde suivie d'une postfixation met en évidence six catégories cellulaires.Les résultats différents obtenus avec ces deux fixateurs portent essentiellement sur la structure (forme et densité) des granulations; une meilleure conservation des grains de sécrétion est obtenue avec la glutaraldéhyde.La libération et l'élaboration des produits de sécrétion sont discutées. Le problème de la libération des granules des cellules hypophysaires a été abordé sous un angle nouveau. Il a été observé dans d'autres organes (pancréas endocrine, cellule à gastrine de l'intestin) que les grains de sécrétion subissent une lyse intracytoplasmique sans destruction de la membrane limitante du granule. Il est suggéré que le même processus pourrait aussi se réaliser au niveau de l'hypophyse.

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On the ultrastructure of the adenohypophysis in the toad, Bufo bufo L.I. Identification of cell types and comparison of the results obtained with different fixations

Summary Two different fixatives have been used to identify the various cell types contained in the anterior pituitary of the toad (Bufo bufo L.) and to investigate their ultrastructure. Five cell types were distinguished after simple osmium tetroxid fixation, whilst glutaraldehyde fixation with postosmification revealed six distinct cell types.The results obtained by these two techniques differed mainly in the structure (shape and electrondensity) of the secretory granules, which seemed to be better preserved after combined glutaraldehyde-osmium fixation.The modes of synthesis and release of the secretory products are discussed. The problem of the mode of release of secretory granules from pituitary cells has been envisaged under a new aspect. It has been observed in other organs (endocrine pancreas, gastrin cells of the intestinal wall) that secretory granules may undergo intracytoplasmic lysis without destruction of their limiting membrane. It is suggested that the same process may operate in the pituitary.

Travail réalisé avec l'aide du Fonds national suisse de la Recherche scientifique (Crédit No 5344.3).Nous remercions tout particulièrement Mme Sidler-Ansermet, M. Oberson, photographes, et Mme Baumann, secrétaire, de l'aide qu'ils ont apportée à la réalisation de ce mémoire.     

Membrane junctions between salivary gland cells ofChironomus thummi

Summary Cells ofChironomus salivary glands communicate through intercellular connections of high permeability. Electron micrographs of salivary glands show two kinds of junctions between the membranes of adjacent cells, which may be responsible for cell coupling: septate junctions and close membrane junctions.A large fraction of lateral cell surfaces is occupied by septate junctions, while the area of close membrane junctions appears to be very small. Consequently septate junctions have been considered as likely sites for intercellular coupling. There are however some indications that intercellular communication is provided by structures which seem to be unstable. As osmotic effects are among the factors which can disrupt cellular communications, we have tried to eliminate possible effects of the fixing solutions on the ultrastructure of intercellular connections by using isoosmotic fixatives. Under these conditions large regions of close membrane junctions of the nexus kind have been observed to occur between gland cells. They are of similar size as septate junctions. It seems to be possible that as in other communicating cell systems nexus could be the sites for intercellular coupling of salivary gland cells.The authors would like to thank Prof. Dr. H. Leonhardt, Institut für Anatomie I, Homburg, for the use of his electron microscope (Zeiss EM 9-DFG grant LE 69–8) during part of this work and Prof. Dr. H. Kroeger, Institut für Genetik, Saarbrücken for the supply withChironomus larvae.     

不同固定方法对植物细胞扫描电镜观察用冷冻断裂样品的影响

用冷冻断裂法在扫描电镜下研究了洋葱(Allium cepa)根端分生组织细胞内部的三维结构。采用了两种固定方法。冷冻断裂前只用1％锇酸固定的材料容易在细胞质和核之间断开,而用卡诺固定液(无水乙醇:冰醋酸3:1)前固定,然后再用1％锇酸固定的材料容易使细胞核断裂。前一固定方法适于研究细胞质的内部结构(细胞骨架的纤维、线粒体、内质网等及其三维分布关系):后一固定方法适于研究核内结构(染色质、核仁、核基质纤维)的三维形象,特别是核仁纤维中心染色质的三维结构。