Immunogold labeling in scanning electron microscopy

1996 ◽  
Vol 106 (1) ◽  
pp. 31-39 ◽  
Author(s):  
R. Hermann ◽  
P. Walther ◽  
M. Müller
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Immunogold Labeling ◽  
Scanning Electron

scholarly journals Immunogold Labeling of Rubisco In C4 Plant Leaves for Scanning Electron Microscopy

2001 ◽  
Vol 4 (1) ◽  
pp. 41-49 ◽  
Author(s):  
Hiroshi Miyake ◽  
Mariko Nishimura ◽  
Yoji Takeoka
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Immunogold Labeling ◽  
Plant Leaves ◽  
C4 Plant ◽  
Scanning Electron

scholarly journals Electron Microscopy Techniques for Investigating Structure and Composition of Hair-Cell Stereociliary Bundles

2021 ◽  
Vol 9 ◽  
Author(s):  
Maryna V. Ivanchenko ◽  
Artur A. Indzhykulian ◽  
David P. Corey
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Sample Preparation ◽  
Protein Localization ◽  
Imaging Techniques ◽  
Ion Beam ◽  
Immunogold Labeling ◽  
Sensory Cells ◽  
Optimal Sample ◽  
Scanning Electron

Hair cells—the sensory cells of the vertebrate inner ear—bear at their apical surfaces a bundle of actin-filled protrusions called stereocilia, which mediate the cells’ mechanosensitivity. Hereditary deafness is often associated with morphological disorganization of stereocilia bundles, with the absence or mislocalization within stereocilia of specific proteins. Thus, stereocilia bundles are closely examined to understand most animal models of hereditary hearing loss. Because stereocilia have a diameter less than a wavelength of light, light microscopy is not adequate to reveal subtle changes in morphology or protein localization. Instead, electron microscopy (EM) has proven essential for understanding stereocilia bundle development, maintenance, normal function, and dysfunction in disease. Here we review a set of EM imaging techniques commonly used to study stereocilia, including optimal sample preparation and best imaging practices. These include conventional and immunogold transmission electron microscopy (TEM) and scanning electron microscopy (SEM), as well as focused-ion-beam scanning electron microscopy (FIB-SEM), which enables 3-D serial reconstruction of resin-embedded biological structures at a resolution of a few nanometers. Parameters for optimal sample preparation, fixation, immunogold labeling, metal coating and imaging are discussed. Special attention is given to protein localization in stereocilia using immunogold labeling. Finally, we describe the advantages and limitations of these EM techniques and their suitability for different types of studies.

scholarly journals Scanning electron microscopy of type I collagen at the dentin-enamel junction of human teeth.

1993 ◽  
Vol 41 (3) ◽  
pp. 381-388 ◽  
Author(s):  
C P Lin ◽  
W H Douglas ◽  
S L Erlandsen
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Type I Collagen ◽  
Immunogold Labeling ◽  
Matrix Composition ◽  
Type I ◽  
Human Teeth ◽  
Dentin Matrix ◽  
Electron Imaging ◽  
Scanning Electron

The dentin-enamel junction constitutes a unique boundary between two highly mineralized tissues with very different matrix composition and physical properties. The nature of the boundary between the ectoderm-derived enamel and mesoderm-derived dentin is not known. This study was undertaken to identify the presence, type, and distribution of collagen at the dentin-enamel junction as an initial step in understanding its structural-functional role in dental occlusion. Sections of human teeth were demineralized with 0.1 M neutral EDTA and examined by high-resolution field-emission scanning electron microscopy at low accelerating voltage. Enamel and dentin were observed to be linked by many parallel 80-120-nm diameter fibrils, which were inserted directly into the enamel mineral and also merged with the interwoven fibrillar network of the dentin matrix. Immunogold labeling for collagen was visualized by secondary electron imaging and backscatter electron imaging at low accelerating voltage. The collagen fibrils at the junctional zone as well as in the dentin matrix were identified as Type I collagen. Collagenase digestion led to loss of the fibrillar structures and prevented immunogold labeling with antibody specific to Type I collagen. Consequently, the dentin-enamel junction can be regarded as a fibril-reinforced bond which is mineralized to a moderate degree.

scholarly journals Morphological analysis of Apolipoprotein E binding to Aβ Amyloid using a combination of Surface Plasmon Resonance, Immunogold Labeling and Scanning Electron Microscopy

2019 ◽  
Vol 19 (1) ◽  
Author(s):  
Tohidul Islam ◽  
Anna L. Gharibyan ◽  
Cheng Choo Lee ◽  
Anders Olofsson
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Surface Plasmon Resonance ◽  
Apolipoprotein E ◽  
Surface Plasmon ◽  
Plasmon Resonance ◽  
Amyloid Β ◽  
Immunogold Labeling ◽  
Aβ Amyloid ◽  
Scanning Electron

Abstract Background Immunogold labeling in combination with transmission electron microscopy analysis is a technique frequently used to correlate high-resolution morphology studies with detailed information regarding localization of specific antigens. Although powerful, the methodology has limitations and it is frequently difficult to acquire a stringent system where unspecific low-affinity interactions are removed prior to analysis. Results We here describe a combinatorial strategy where surface plasmon resonance and immunogold labeling are used followed by a direct analysis of the sensor-chip surface by scanning electron microscopy. Using this approach, we have probed the interaction between amyloid-β fibrils, associated to Alzheimer’s disease, and apolipoprotein E, a well-known ligand frequently found co-deposited to the fibrillar form of Aβ in vivo. The results display a lateral binding of ApoE along the amyloid fibrils and illustrates how the gold-beads represent a good reporter of the binding. Conclusions This approach exposes a technique with generic features which enables both a quantitative and a morphological evaluation of a ligand-receptor based system. The methodology mediates an advantage compared to traditional immunogold labeling since all washing steps can be monitored and where a high stringency can be maintained throughout the experiment.

scholarly journals Immunogold labeling in scanning electron microscopy

1996 ◽  
Vol 106 (3) ◽  
pp. 356-356 ◽  
Author(s):  
R. Hermann ◽  
P. Walther ◽  
M. Müller
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Immunogold Labeling ◽  
Scanning Electron

Immunogold labeling in scanning electron microscopy

1996 ◽  
Vol 106 (1) ◽  
pp. 31-39 ◽  
Author(s):  
René Hermann ◽  
Paul Walther ◽  
M. Müller
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Immunogold Labeling ◽  
Scanning Electron

scholarly journals Immunogold Labeling of Amelogenin in Developing Porcine Enamel Revealed by Field Emission Scanning Electron Microscopy

2008 ◽  
Vol 189 (1-4) ◽  
pp. 207-211 ◽  
Author(s):  
Chang Du ◽  
Daming Fan ◽  
Zhi Sun ◽  
Yuwei Fan ◽  
Rajamani Lakshminarayanan ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Field Emission ◽  
Immunogold Labeling ◽  
Scanning Electron

Pathogenesis of Human Hair Defects

1974 ◽  
Vol 32 ◽  
pp. 12-13
Author(s):  
P.S. Porter ◽  
T. Aoyagi ◽  
R. Matta
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Human Hair ◽  
Standard Techniques ◽  
Scanning Electron

Using standard techniques of scanning electron microscopy (SEM), over 1000 human hair defects have been studied. In several of the defects, the pathogenesis of the abnormality has been clarified using these techniques. It is the purpose of this paper to present several distinct morphologic abnormalities of hair and to discuss their pathogenesis as elucidated through techniques of scanning electron microscopy.

Ultrastructural Analysis of the Salivary Glands of Gromphadorhina Portentosa (Schaum)

1977 ◽  
Vol 35 ◽  
pp. 638-639
Author(s):  
P.J. Dailey
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Salivary Glands ◽  
Secretory Vesicles ◽  
The Past ◽  
Transmission Electron ◽  
Means Of Transmission ◽  
Gromphadorhina Portentosa ◽  
Scanning Electron ◽  
Topographical Relief

The structure of insect salivary glands has been extensively investigated during the past decade; however, none have attempted scanning electron microscopy (SEM) in ultrastructural examinations of these secretory organs. This study correlates fine structure by means of SEM cryofractography with that of thin-sectioned epoxy embedded material observed by means of transmission electron microscopy (TEM).Salivary glands of Gromphadorhina portentosa were excised and immediately submerged in cold (4°C) paraformaldehyde-glutaraldehyde fixative1 for 2 hr, washed and post-fixed in 1 per cent 0s04 in phosphosphate buffer (4°C for 2 hr). After ethanolic dehydration half of the samples were embedded in Epon 812 for TEM and half cryofractured and subsequently critical point dried for SEM. Dried specimens were mounted on aluminum stubs and coated with approximately 150 Å of gold in a cold sputtering apparatus.Figure 1 shows a cryofractured plane through a salivary acinus revealing topographical relief of secretory vesicles.

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