New polarized-light microscope for fast and orientation independent measurement of birefringent fine structure

Author(s):  
Rudolf Oldenbourg

The polarized light microscope has the unique potential to measure submicroscopic molecular arrangements dynamically and non-destructively in living cells and other specimens. With the traditional pol-scope, however, single images display only those anisotropic structures that have a limited range of orientations with respect to the polarization axes of the microscope. Furthermore, rapid measurements are restricted to a single image point or single area that exhibits uniform birefringence or other form of optical anisotropy, while measurements comparing several image points take an inordinately long time.We are developing a new kind of polarized light microscope which combines speed and high resolution in its measurement of the specimen anisotropy, irrespective of its orientation. The design of the new pol-scope is based on the traditional polarized light microscope with two essential modifications: circular polarizers replace linear polarizers and two electro-optical modulators replace the traditional compensator. A video camera and computer assisted image analysis provide measurements of specimen anisotropy in rapid succession for all points of the image comprising the field of view.

Author(s):  
Rudolf Oldenbourg

The recent renaissance of the light microsope is fueled in part by technological advances in components on the periphery of the microscope, such as the laser as illumination source, electronic image recording (video), computer assisted image analysis and the biochemistry of fluorescent dyes for labeling specimens. After great progress in these peripheral parts, it seems timely to examine the optics itself and ask how progress in the periphery facilitates the use of new optical components and of new optical designs inside the microscope. Some results of this fruitful reflection are presented in this symposium.We have considered the polarized light microscope, and developed a design that replaces the traditional compensator, typically a birefringent crystal plate, with a precision universal compensator made of two liquid crystal variable retarders. A video camera and digital image processing system provide fast measurements of specimen anisotropy (retardance magnitude and azimuth) at ALL POINTS of the image forming the field of view. The images document fine structural and molecular organization within a thin optical section of the specimen.


2011 ◽  
Vol 402 ◽  
pp. 407-411 ◽  
Author(s):  
Jacob M. Mchenya ◽  
Sheng Zhuo Zhang ◽  
Song Jing Li

In order to understand the mechanism and get rid of the high-frequency self-excited noise in a hydraulic servo-valve, in this paper, the flow field distribution in the pilot stage of a hydraulic flapper-nozzle servo-valve is investigated. An assembly is prepared representing the construction and working principle of the flow field inside the pilot stage of a hydraulic flapper-nozzle servo-valve. A method of visualization is developed by taking videos for the flow field inside the transparent assembly with a high speed video camera. In this study, at different inlet pressure the high speed video camera was utilized for flow visualization together with computer-assisted image measurement. The shape of the jet flow, the cavitations and vortex flow inside the flow field can be visualized clearly. The proposed method enables to analyze the flow-field in the pilot stage of a hydraulic flapper-nozzle servo-valve by giving useful information for better design.


Author(s):  
A.M. Jones ◽  
A. Max Fiskin

If the tilt of a specimen can be varied either by the strategy of observing identical particles orientated randomly or by use of a eucentric goniometer stage, three dimensional reconstruction procedures are available (l). If the specimens, such as small protein aggregates, lack periodicity, direct space methods compete favorably in ease of implementation with reconstruction by the Fourier (transform) space approach (2). Regardless of method, reconstruction is possible because useful specimen thicknesses are always much less than the depth of field in an electron microscope. Thus electron images record the amount of stain in columns of the object normal to the recording plates. For single particles, practical considerations dictate that the specimen be tilted precisely about a single axis. In so doing a reconstructed image is achieved serially from two-dimensional sections which in turn are generated by a series of back-to-front lines of projection data.


Author(s):  
Walter C. McCrone

An excellent chapter on this subject by V.D. Fréchette appeared in a book edited by L.L. Hench and R.W. Gould in 1971 (1). That chapter with the references cited there provides a very complete coverage of the subject. I will add a more complete coverage of an important polarized light microscope (PLM) technique developed more recently (2). Dispersion staining is based on refractive index and its variation with wavelength (dispersion of index). A particle of, say almandite, a garnet, has refractive indices of nF = 1.789 nm, nD = 1.780 nm and nC = 1.775 nm. A Cargille refractive index liquid having nD = 1.780 nm will have nF = 1.810 and nC = 1.768 nm. Almandite grains will disappear in that liquid when observed with a beam of 589 nm light (D-line), but it will have a lower refractive index than that liquid with 486 nm light (F-line), and a higher index than that liquid with 656 nm light (C-line).


Author(s):  
F.A. Ponce ◽  
H. Hikashi

The determination of the atomic positions from HRTEM micrographs is only possible if the optical parameters are known to a certain accuracy, and reliable through-focus series are available to match the experimental images with calculated images of possible atomic models. The main limitation in interpreting images at the atomic level is the knowledge of the optical parameters such as beam alignment, astigmatism correction and defocus value. Under ordinary conditions, the uncertainty in these values is sufficiently large to prevent the accurate determination of the atomic positions. Therefore, in order to achieve the resolution power of the microscope (under 0.2nm) it is necessary to take extraordinary measures. The use of on line computers has been proposed [e.g.: 2-5] and used with certain amount of success.We have built a system that can perform operations in the range of one frame stored and analyzed per second. A schematic diagram of the system is shown in figure 1. A JEOL 4000EX microscope equipped with an external computer interface is directly linked to a SUN-3 computer. All electrical parameters in the microscope can be changed via this interface by the use of a set of commands. The image is received from a video camera. A commercial image processor improves the signal-to-noise ratio by recursively averaging with a time constant, usually set at 0.25 sec. The computer software is based on a multi-window system and is entirely mouse-driven. All operations can be performed by clicking the mouse on the appropiate windows and buttons. This capability leads to extreme friendliness, ease of operation, and high operator speeds. Image analysis can be done in various ways. Here, we have measured the image contrast and used it to optimize certain parameters. The system is designed to have instant access to: (a) x- and y- alignment coils, (b) x- and y- astigmatism correction coils, and (c) objective lens current. The algorithm is shown in figure 2. Figure 3 shows an example taken from a thin CdTe crystal. The image contrast is displayed for changing objective lens current (defocus value). The display is calibrated in angstroms. Images are stored on the disk and are accessible by clicking the data points in the graph. Some of the frame-store images are displayed in Fig. 4.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Wanrong Gao

AbstractIn this work, we introduce the concept of anisotropic dielectric susceptibility matrix of anisotropic medium for both nondepolarizing and depolarizing medium. The concept provides a new way of analyzing light scattering properties of anisotropic media illuminated by polarized light. The explicit expressions for the elements of the scattering matrix are given in terms of the elements of the Fourier transform of the anisotropic dielectric susceptibility matrix of the medium. Finally, expressions for the elements of the Jones matrix of a thin layer of a deterministic anisotropic medium and the elements of the Mueller matrix of a depolarizing medium are given. The results obtained in this work is helpful for deriving information about the correlated anisotropic structures in depolarizing media from measured Mueller matrices. The findings in this work may also well prove stimulating to researchers working on new methods for analyzing light scattering properties.


2019 ◽  
Vol 6 ◽  
pp. 1-8 ◽  
Author(s):  
Marie Beermann ◽  
Johan Lindeberg ◽  
Jennie Engstrand ◽  
Karolina Galmén ◽  
Silja Karlgren ◽  
...  

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