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Statistical analysis of selected parameters of biological cells investigated with digital holographic microscopy

Patryk Kamil Nienałtowski

Abstract

The motivation to implement this engineering thesis is the progressive development of quantitative phase imaging methods in biomedical applications, as well as the need for statistical analysis of the results obtained in these methods. In particular I have focused on the statistical analysis of the phase results of cell cultures obtained in the digital holographic microscope (DHM). The uniqueness of DHM consists in capability to analyze transparent or quasitransparent cells in the label-free (no interference of markers with biomaterial) and quantitative manner. Due to this feature digital holographic microscopy can become the basic tool for cell and tissue investigations and in the future even replace traditional optical imaging. The aim of the engineering thesis is to develop a methodology and tools for statistical analysis of selected cell parameters based on phase measurements. The thesis contains the review of available solutions for the analysis of biological images and the assessment of their utility for phase images analysis. Due to numerous disadvantages of the available solutions and their incongruity to DIC requirements I had decided that a new software for image processing and analysis of an integrated phase should be developed. The algorithms should ensure proper image preprocessing and segmentation, individual cells separation and determination of their individual parameters. The software developed in the thesis was tested on two examples of analysis of DIC results, namely investigations of the cultures of: NIH3T3 fibroblasts and HaCaT keratinocytes. The results for fibroblast analysis were obtained by MSc M. Baczewska in the laboratory at the Westfalen University of Wilhelm in Münster. HaCaT keratinocyte cultures were prepared by a team from the Medical University of Warsaw and I carried out the holographic measurements of cells during their apoptosis process in the laboratory of the Institute of Micromechanics and Photonics, WUT. Both measuring systems were based on the Mach-Zehnder interferometer architecture with a camera capturing the off-axis image plane holograms. This allowed to use in both cases the simplified method of phase reconstruction by the Fourier Transform Method. The comparison of statistical results of the analysis performed at live and fixed NIH3T3 fibroblasts has showed clear differences in the values of dry mass and phase volume for the significance level of 6%. It was also found that the surfaces of live and fixed fibroblasts do not differ from each other with a 3% probability of error. In the case of HaCaT keratinocyte series, the ability to monitor and analyze different stages of cell life based on the parameters correlated with measured phases was demonstrated. The tools developed during this thesis and the obtained results create good basis for introducing a digital holographic microscope as a quantitative tool for full and reliable analysis of biophysical parameters of biological material.
Diploma type
Engineer's / Bachelor of Science
Diploma type
Engineer's thesis
Author
Patryk Kamil Nienałtowski (FM) Patryk Kamil Nienałtowski,, Faculty of Mechatronics (FM)
Title in Polish
Analiza statystyczna wybranych parametrów komórek biologicznych zmierzonych w układzie cyfrowego mikroskopu holograficznego
Supervisor
Małgorzata Kujawińska (FM/IMPh) Małgorzata Kujawińska,, The Institute of Micromechanics and Photonics (FM/IMPh)Faculty of Mechatronics (FM)
Certifying unit
Faculty of Mechatronics (FM)
Affiliation unit
The Institute of Micromechanics and Photonics (FM/IMPh)
Study subject / specialization
, Mechatronika
Language
(pl) Polish
Status
Finished
Defense Date
07-02-2019
Issue date (year)
2019
Reviewers
Małgorzata Kujawińska (FM/IMPh) Małgorzata Kujawińska,, The Institute of Micromechanics and Photonics (FM/IMPh)Faculty of Mechatronics (FM) Wojciech Krauze (FM/IMPh) Wojciech Krauze,, The Institute of Micromechanics and Photonics (FM/IMPh)Faculty of Mechatronics (FM)
Keywords in Polish
fazowe obrazowanie ilościowe, cyfrowa mikroskopia holograficzna, segmentacja obrazów biologicznych, sucha masa, fibroblasty NIH3T3, keratynocyty HaCaT, analiza statystyczna.
Keywords in English
quantitative phase imaging, digital holographic microscopy, biological image segmentation, dry mass, NIH3T3 fibroblasts, HaCaT keratinocytes, statistical analysis
Abstract in Polish
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Identyfikator pracy APD: 31536

Uniform Resource Identifier
https://repo.pw.edu.pl/info/bachelor/WUT87dfea914fe14c11a57e322cf23cebc4/
URN
urn:pw-repo:WUT87dfea914fe14c11a57e322cf23cebc4

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