A Novel, Label-Free Method for Quantifying Collagen Fibril Formation and Degradation Using DIC Microscopy, Electromagnetic Wave Theory, and Image Processing

Author(s):  
Brendan P. Flynn ◽  
Amit Bhole ◽  
Charles DiMarzio ◽  
Jeffrey W. Ruberti

Methods to assay fibrillar growth and degradation at sub-light scales include: fluorescence assays using FITC-collagen or FRAP, destructive preparation and measurement using electron microscopy, and light occlusion methods including turbidity and absorption methods. Many of these methods require the outright destruction, or at least modification via labelling, of the sample in question. This requirement can slow experimentation and introduce additional variability or even alter the reaction rate kinetics. The two methods (absorption and turbidity) which are label-free are bulk averaging methods and cannot isolate subsets of fibrils (e.g. fibrils under load).

Author(s):  
R.W. Horne

The technique of surrounding virus particles with a neutralised electron dense stain was described at the Fourth International Congress on Electron Microscopy, Berlin 1958 (see Home & Brenner, 1960, p. 625). For many years the negative staining technique in one form or another, has been applied to a wide range of biological materials. However, the full potential of the method has only recently been explored following the development and applications of optical diffraction and computer image analytical techniques to electron micrographs (cf. De Hosier & Klug, 1968; Markham 1968; Crowther et al., 1970; Home & Markham, 1973; Klug & Berger, 1974; Crowther & Klug, 1975). These image processing procedures have allowed a more precise and quantitative approach to be made concerning the interpretation, measurement and reconstruction of repeating features in certain biological systems.


Author(s):  
Yasushi Kokubo ◽  
Hirotami Koike ◽  
Teruo Someya

One of the advantages of scanning electron microscopy is the capability for processing the image contrast, i.e., the image processing technique. Crewe et al were the first to apply this technique to a field emission scanning microscope and show images of individual atoms. They obtained a contrast which depended exclusively on the atomic numbers of specimen elements (Zcontrast), by displaying the images treated with the intensity ratio of elastically scattered to inelastically scattered electrons. The elastic scattering electrons were extracted by a solid detector and inelastic scattering electrons by an energy analyzer. We noted, however, that there is a possibility of the same contrast being obtained only by using an annular-type solid detector consisting of multiple concentric detector elements.


Author(s):  
J. H. Hayden

In a previous study, Allen video-enhanced constrast/differential interference constrast (AVEC-DIC) microscopy was used in conjunction with immunofluorescence microscopy to demonstrate that organelles and vesicle move in either direction along linear elements composed of microtubules. However, this study was limited in that the number of microtubules making up a linear element could not be determined. To overcome this limitation, we have used AVEC-DIC microscopy in conjunction with whole mount electron microscopy.Keratocytes from Rana pipiens were grown on glass coverslips as described elsewhere. Gold London Finder grids were Formvar- and carbon coated, and sterilized by exposure to ultraviolet light. It is important to select a Formvar film that gives a grey reflection when it is floated on water. A silver film is too thick and will detract from the image in the light microscope.


2019 ◽  
Vol 29 (1) ◽  
pp. 1226-1234
Author(s):  
Safa Jida ◽  
Hassan Ouallal ◽  
Brahim Aksasse ◽  
Mohammed Ouanan ◽  
Mohamed El Amraoui ◽  
...  

Abstract This work intends to apprehend and emphasize the contribution of image-processing techniques and computer vision in the treatment of clay-based material known in Meknes region. One of the various characteristics used to describe clay in a qualitative manner is porosity, as it is considered one of the properties that with “kill or cure” effectiveness. For this purpose, we use scanning electron microscopy images, as they are considered the most powerful tool for characterising the quality of the microscopic pore structure of porous materials. We present various existing methods of segmentation, as we are interested only in pore regions. The results show good matching between physical estimation and Voronoi diagram-based porosity estimation.


2021 ◽  
Vol 27 (S1) ◽  
pp. 94-95
Author(s):  
Ryan Lane ◽  
Luuk Balkenende ◽  
Simon van Staalduine ◽  
Anouk Wolters ◽  
Ben Giepmans ◽  
...  

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