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
1996 ◽  
Vol 106 (1) ◽  
pp. 31-39 ◽  
Author(s):  
R. Hermann ◽  
P. Walther ◽  
M. Müller

Author(s):  
Maryna V. Ivanchenko ◽  
Artur A. Indzhykulian ◽  
David P. Corey

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.


1993 ◽  
Vol 41 (3) ◽  
pp. 381-388 ◽  
Author(s):  
C P Lin ◽  
W H Douglas ◽  
S L Erlandsen

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.


2019 ◽  
Vol 19 (1) ◽  
Author(s):  
Tohidul Islam ◽  
Anna L. Gharibyan ◽  
Cheng Choo Lee ◽  
Anders Olofsson

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.


1996 ◽  
Vol 106 (3) ◽  
pp. 356-356 ◽  
Author(s):  
R. Hermann ◽  
P. Walther ◽  
M. Müller

1996 ◽  
Vol 106 (1) ◽  
pp. 31-39 ◽  
Author(s):  
René Hermann ◽  
Paul Walther ◽  
M. Müller

2008 ◽  
Vol 189 (1-4) ◽  
pp. 207-211 ◽  
Author(s):  
Chang Du ◽  
Daming Fan ◽  
Zhi Sun ◽  
Yuwei Fan ◽  
Rajamani Lakshminarayanan ◽  
...  

2014 ◽  
Vol 900 ◽  
pp. 289-292
Author(s):  
Qing Cheng Wang ◽  
Xiao Dong Yang ◽  
Guang Rui Shang

This paper studied wettability of several typical plant leaves such as abutilon, Jerusalem artichoke and sunflower, contact angle of several leaves were measured through the contact angle measuring instrument, which were 41.5°, 46.3°and 43.6°respectively. Microstructure of abutilon, Jerusalem artichoke and sunflower leaves were observed by scanning electron microscopy, It found that several typical plant leaves are composite structure of micron scale and nanoscale, covered some long needle-like spikes, this feature has an important influence on hydrophilicity. According to wenzel model, the paper analyzed the relationship between hydrophilicity and microstructures of several plant leaves.


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