Multicolor Fluorescence Microscopy Using the Laser-Scanning Confocal Microscope
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Microstructure codes for the properties of food. Processing enables the microstructure. Food microstructures are in most cases hierarchical, heterogeneous, multiphase, and complex. A full understanding of the food microstructure requires the characterization at many different length scales. Light microscopy and confocal laser scanning microscopy are powerful tools to image food microstructures at the micrometer level. In this chapter, the principles and use of these microscopy techniques are described. Examples of the use of light microscopy and confocal laser scanning microscopy to characterize and understand the microstructures in bread and dough, fibrous vegetable protein structures, plant cell walls, fat-rich food, and mayonnaise are discussed. In the end, an outlook on the use of light microscopy and confocal laser scanning microscopy in foods is given.
Laser Microscopy
Characterization
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Two-photon excitation microscopy
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Light microscopy and confocal scanning laser microscopy are widely used imaging techniques to generate structural insights in the complex interplay of components in food and processed food. Focusing on the application of optical microscopy (light and confocal scanning laser microscopy) to dairy products, this chapter briefly discusses the history of the technique and evolution of food microscopy. It describes and discusses the differences in imaging modes as well as the methodology to obtain a specimen or sub-sample that represents the original and undisturbed microstructure. Staining and specific labeling procedures are discussed for both bright field and confocal laser scanning microscopy. It reflects on applications for dynamic imaging using temperature control and shear stages to study microstructural changes during processing.
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We propose a framework to quantify photodamage in multiphoton light-sheet microscopy. Using cardiac imaging in live zebrafish embryos, we demonstrate an order of magnitude signal enhancement is safely obtained by adjusting the laser repetition rate.
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We propose a framework to quantify photodamage in multiphoton light-sheet microscopy. Using cardiac imaging in live zebrafish embryos, we demonstrate an order of magnitude signal enhancement is safely obtained by adjusting the laser repetition rate.
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Laser light
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INTRODUCTION Fluorescence microscopy has been gaining importance in quantitative biological research due to dramatic improvements in fluorophores, optical systems, light sources, and detectors. In particular, confocal fluorescence microscopy, usually by laser scanning, has for the first time allowed the observation of biological processes with high spatial resolution inside intact living tissue. Often called “optical sectioning,” this method allows spatial reconstruction of 3D specimens without the use of a microtome. This article presents the physical mechanisms upon which the properties of multiphoton microscopy are based and discusses some practical aspects of its implementation.
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This paper introduces the development of the hard cholecystoscope of confocal laser scanning,which combined the confocal laser scanning system and the hard cholecystoscope.The hard cholecystoscope of confocal laser scanning can do the laser scanning when the cells are alive,and can provide the microcosmic images with the gallbladder's pathological changes.By comparing these images,the doctors can make the precise diagnosis.
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This chapter focuses on the basics of the light sheet microscopy principle and construction, followed by an overview of its various implementations. It discusses some of the novel data handling solutions that light sheet microscopy gave rise to, as well as future prospects in this area. Two different ways of generating a light sheet are commonly used: in selective plane illumination microscopy (SPIM), the laser beam is expanded and focused using a cylindrical lens to form the sheet of light. Alternatively, a "virtual" light sheet can be generated by the digitally scanned light sheet microscopy (DSLM) technique, which uses a scanning mirror to rapidly sweep a beam across the field of view (FOV). The chapter discusses how to acquire 2D images of a sample using light sheet microscopy. However, most biological applications require 3D imaging. Given its excellent optical sectioning capability, speed, and low phototoxicity, light sheet microscopy is ideally suited for fast 3D imaging.
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