Electron Microscopic in Cellular and Molecular Biology

Plattner, Helmut; Zingsheim, H. P.

doi:10.1007/978-1-4613-3533-7_1

Cited by 33 publications

(35 citation statements)

References 678 publications

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“…Postembedding Au-labeled EM sections [fixation (b)] were used for quantitation. Gold label was referred to the size of the structures evaluated and this was determined by the hit point method (see Plattner and Zingsheim 1983).…”

Section: Postembedding Immunogold Labelingmentioning

confidence: 99%

Immunolocalization of the exocytosis-sensitive phosphoprotein, PP63/parafusin, in Paramecium cells using antibodies against recombinant protein

Kissmehl

Hauser²,

Gößringer³

et al. 1998

Histochemistry and Cell Biology

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We have localized a structure-bound fraction of the exocytosis-sensitive phosphoprotein, PP63/parafusin (PP63/pf), in Paramecium cells by widely different methods. We combined cell fractionation, western blots, as well as light and electron microscopy (pre-and postembedding immunolabeling), applying antibodies against the recombinant protein. PP63/pf is considerably enriched in certain cortical structures, notably the outlines of regular surface fields (kinetids), docking sites of secretory organelles (trichocysts) and the membranes of subplasmalemmal Ca 2+ -stores (alveolar sacs). From our localization studies we tentatively derive several potential functions for PP63/pf, including cell surface structuring, assembly of exocytosis sites, and/or Ca 2+ homeostasis.& b d y :

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Section: Postembedding Immunogold Labelingmentioning

confidence: 99%

Immunolocalization of the exocytosis-sensitive phosphoprotein, PP63/parafusin, in Paramecium cells using antibodies against recombinant protein

Kissmehl

Hauser²,

Gößringer³

et al. 1998

Histochemistry and Cell Biology

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“…Hara & Chihara (1987) reported similar results for the ultrastructural studies of H. akashiwo. Hence, it should be noted that morphological damage might be easily caused by chemical fixation (Plattnerr & Zingsheim 1983).…”

Section: Resultsmentioning

confidence: 99%

“…For example, aldehydes (e.g., glutaraldehyde, formaldehyde) are often used to bridge and link distant pairs of protein molecules in the algal cells. However, this fixation method frequently damages selective membrane permeability, leading to the formation of artifact products and changes to the cell ultrastructure (Plattnerr & Zingsheim 1983).…”

Section: Introductionmentioning

confidence: 99%

Ultrastructural observation of natural field phytoplankton cells by using rapid freezing and freeze substitution

Kimura

Tomaru

Nagasaki

2012

Plankton Benthos Res

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Transmission electron microscopy (TEM) is a basic technique used to study the ultrastructures of phytoplankton. However, the use of chemical fixation to prepare phytoplankton samples for TEM is laborious and timeconsuming. Furthermore, chemical fixation often causes artificial structural changes (artifacts) in phytoplankton cells. In this study, we induced and optimized a rapid freezing and freeze substitution (RFS) method, accompanied by a filter dehydration system, to prepare TEM samples of marine microalgae, including Heterosigma akashiwo, Chattonella antiqua (Raphidophyceae), Heterocapsa circularisquama (Dinophyceae), Chaetoceros tenuissimus (Bacillariophyceae), and Teleaulax amphioxeia (Cryptophyceae). The whole procedure was much more efficient than the traditional method. In addition, the fine structure of each microalgal cell was well preserved. For example, the fine structures of the nuclear membrane, nuclear pores, and several vesicles were clearly observed, which were comparatively difficult to observe using the chemical fixation method. In addition, this new technique was applicable to the natural blooming cells of H. akashiwo, and high-resolution ultrastructural images were successfully observed. This is the first report on the application of the modified-RFS method for natural phytoplankton assemblages. This method can be used for diverse phytoplankters in natural waters, and the high-quality observations are expected to provide useful information toward understanding phytoplankter physiology and ecology.

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“…Considering I SUB-SECOND CELLULAR DYNAMICS the diameter of one transmembrane stretch (Eskandari et al. 1998;Plattner and Zingsheim, 1983) one can estimate the number of molecules required to make up a freeze-fracture particle, such as a rosette particle at a trichocyst docking/fusion site (Plattner and Kissmehl. 2003a).…”

Section: A Membrane Fusion and Fissionmentioning

confidence: 99%

Sub-Second Cellular Dynamics: Time-Resolved Electron Microscopy and Functional Correlation

Plattner¹,

Hentschel²

2006

International Review of Cytology

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Subcellular processes, from molecular events to organellar responses and cell movement, cover a broad scale in time and space. Clearly the extremes, such as ion channel activation are accessible only by electrophysiology, whereas numerous routine methods exist for relatively slow processes. However, many other processes, from a millisecond time scale on, can be "caught" only by methods providing appropriate time resolution. Fast freezing (cryofixation) is the method of choice in that case. In combination with follow-up methodologies appropriate for electron microscopic (EM) analysis, with all its variations, such technologies can also provide high spatial resolution. Such analyses may include, for example, freeze-fracturing for analyzino restructuring of membrane components, scanning EM and other standard EM techniques, as well as analytical EM analyses. The latter encompass energy-dispersive x-ray microanalysis and electron spectroscopic imaging, all applicable, for instance, to the second messenger, calcium. Most importantly, when conducted in parallel, such analyses can provide a structural background to the functional analyses, such as cyclic nucleotide formation or protein de-or rephosphorylation during cell stimulation. In sum, we discuss many examples of how it is practically possible to achieve strict function-structure correlations in the sub-second time range. We complement this review by discussing alternative methods currently available to analyze fast cellular phenomena occurring in the sub-second time range.

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Electron Microscopic in Cellular and Molecular Biology

Cited by 33 publications

References 678 publications

Immunolocalization of the exocytosis-sensitive phosphoprotein, PP63/parafusin, in Paramecium cells using antibodies against recombinant protein

Immunolocalization of the exocytosis-sensitive phosphoprotein, PP63/parafusin, in Paramecium cells using antibodies against recombinant protein

Ultrastructural observation of natural field phytoplankton cells by using rapid freezing and freeze substitution

Sub-Second Cellular Dynamics: Time-Resolved Electron Microscopy and Functional Correlation

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