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Methodology of Dictyostelium discoideum imaging with the transmission digital holographic microscope
Křížová, Aneta ; Ježek, Jan (referee) ; Uhlířová, Hana (advisor)
The task of this bachelor’s thesis was to image Dictyostelium discoideum with a transmission digital holographic microscope and compare this imaging with a commonly used microscopy technique. This bachelor's thesis is structured into a theoretical part and an experimental part. The theoretical part describes the setting of the transmission digital holographic microscope, basic operation and physical principles of imaging with this microscope. Further the process of image reconstruction and numerical process of phase reconstruction is illustrated. At the end of this part, basic biological characteristics of observed cells of Dictyostelium discoideum are introduced. The experimental part describes the cultivation and preparation of Dictyostelium discoideum for the observations, and further the process of performed experiments. The cells of Dictyostelium discoideum have been imaged in couple experiments by a method of the classical phase contrast. By the transmission digital holographic microscope the cells of Dictyostelium discoideum have been imaged in two mediums. Images captured by the transmission digital holographic microscope were processed by a method of Dynamic phase differences and analysed.
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User programme for imaging and visualization of quantitative phase from the digital holographic microscope
Hladík, Lukáš ; Kvasnica, Lukáš (referee) ; Uhlířová, Hana (advisor)
This bachelor thesis deals with creation of an user computer program for processing of phase contrast images from a digital holographic microscope. The theoretical section of the thesis contains basic information about obtaining and evaluation of processed images, including a description of their digital representation in the computer. The experimental part deals with techniques and algorithms on which the program is built. This part also contains a description of the graphical user interface of the program and evaluate the impact of different user settings for output files properties.
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Correction of phase image background in the digital holographic microscope using convolution
Rudolfová, Zdena ; Malina, Radomír (referee) ; Uhlířová, Hana (advisor)
This bachelor thesis concentrates on a correction of background in the phase image of cells from a transmitted-light digital holographic microscope (TDHM). Optical aberrations in the optical systems of the object and reference arms cause "deformation" of the phase image. The cell is not displayed on a background with a constant value of phase, it is displayed on a background with phase values continuously changing between different places in the image. A method for removing the consequence of this imperfection from the image is described in this thesis. The principle lies in partial convolution of the phase image with a convolution kernel determined by a two-dimensional Gaussian function with the same variance in the both axes. To eliminate loss of information about the cells, the convolution is computed only from those parts of the phase image which contain no cells. The result of the convolution is considered to be an approximation of the background. This approximation is then subtracted from the phase image. The resulting phase image has a constant background, containing only noise. The necessary mathematical concepts are resumed in the introductory part of the thesis. The basic principle of the TDHM image reconstruction is also described. As a part of the thesis, a computer programme Odečet pozadí (Background subtraction) was created, which processes the phase images from the TDHM using this method.
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