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Biophysical interpretation of quantitative phase imaging of live cells generated by coherence-controlled holographic microscopy
Šuráňová, Markéta ; Rösel,, Daniel (referee) ; Vomastek, Tomáš (referee) ; Veselý, Pavel (advisor)
The dissertation thesis deals with the biophysical interpretation of quantitative phase imaging (QPI – Quantitative Phase Imaging) obtained using coherence-controlled holographic microscopy (CCHM – Coherence-Controlled Holographic Microscopy) in the Q-PHASE microscope, Telight, Brno). The theoretical part of this thesis deals with the characteristics of quantitative phase imaging, which provides non-invasive information on the activity of living cells in vitro. The main part of the work consists in elaborating a concept and verifying it of a new methodology (PAMP – Primary Assessment of Migrastatic Potential) for the first critical evaluation of drugs for expected anti-migratory/metastatic potential. The result of this method is considered the first sorting evaluation when considering specific migrastatic agents for future complex oncological treatment. PAMP evaluates the speed of cell migration, the growth of tumor cells and controls the risk of appearance of invasive phenotypes. Furthermore, the correlation microscopy method between the Q-PHASE microscope and the laser scanning confocal microscope (LSCM) is proposed to evaluate cell behavior and the occurrence of focal adhesions after drug application. The quantitative phase image obtained using the Q-PHASE microscope is compared with the quantitative phase image from the HoloMonitor (PHI AB, Sweden), on which the PAMP method has been positively verified.
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Study of the effect of migrastatics on the dynamics of tumour cell migration using a coherence-controlled holographic microscope
Muchová, Nikola ; Netíková,, Irena Štenglová (referee) ; Veselý, Pavel (advisor)
The thesis is focused on the exploitation of the coherence controlled holographic microscopy for the investigation of the influence of the supposed migrastatic drugs on the dynamics of cancer cell migration. The theoretical part briefly describes the history of holographic microscopy and the development of the holographic microscope at the Brno University of Technology in collaboration with Telight, Brno, including an insight into the design and principle of operation of the coherence-controlled holographic microscope. Next a brief description of current cancer treatments introduces new class of anti-cancer drug candidates designated migrastatics that should impair cancer cell migration and thus prevent late metastases formation. The main part of the thesis deals with the design of the experimental observation procedure and data processing using the holographic incoherent Quantitative Phase Imaging technique. The last part of the thesis is the subsequent analysis of the migrastatic effect of the selected drugs on cancer cells in vitro and the comparison of the obtained results with existing studies. Finally, the objectives with magnification 4x and 10x were evaluated and compared for starting “the Rapid Assessment of Cell Growth and Migration”.
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Identification of changes in the behaviour of tumour cells in low concentrations of migrastatics by coherence-controlled holographic microscope
Zábranská, Magdaléna ; Netíková,, Irena Štenglová (referee) ; Veselý, Pavel (advisor)
The subject of the diploma thesis was using of Coherence-Controlled Holographic Microscope in the research of the effect of migrastatics at different concentrations on tumor cells and the interpretation of quantitative phase imaging data. The theoretical introduction is divided into two parts. The first part is devoted to CCHM, which briefly describes the history of holographic microscopy, the principle and a detailed description of the main parts of the microscope. The second part is an introduction to the issue of cancer treatment and the introduction of migrastatic drugs. The experimental part of the work is the analysis of the influence of migrastatics on two tumor lines (H1299 and HT1080) from the measured time-lapse records taken by a holographic microscope. The effect of drugs was evaluated using characteristics describing morphological and dynamic cell changes in the SophiQ software. The data were then graphically and statistically processed to compare and evaluate the effect of migrastatics at low levels. In conclusion of the work, the findings were summarized and selected characteristics were proposed to analyze the impact of migrastatics.
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Analysis of changes in the phenotype of tumour cells induced by migrastatics in quantitative phase imaging
Kolínková, Veronika ; Netíková,, Irena Štenglová (referee) ; Veselý, Pavel (advisor)
This thesis focuses on the observation of living cells using the non-invasive methods of quantitative phase imaging (hiQPI). The imaging is enabled by a coherence-controlled holographic microscope (CCHM) developed in the Laboratory of Experimental Biophotonics at the VUT. Using this imaging technology, morphological changes of A549 and MCF7 after application of potential migrastatic drugs tumor cells are evaluated in the experimental part of the thesis. Migrastatics are defined as already approved drugs that could prevent the migration of cancer cells from the primary tumor and thus prevent metastasis. The RAC-GM (Rapid Assessment of Cell Growth and Migration in Vitro) method was chosen to assess their effect on tumor cells.
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Study of the effect of migrastatics on the dynamics of tumour cell migration using a coherence-controlled holographic microscope
Muchová, Nikola ; Netíková,, Irena Štenglová (referee) ; Veselý, Pavel (advisor)
The thesis is focused on the exploitation of the coherence controlled holographic microscopy for the investigation of the influence of the supposed migrastatic drugs on the dynamics of cancer cell migration. The theoretical part briefly describes the history of holographic microscopy and the development of the holographic microscope at the Brno University of Technology in collaboration with Telight, Brno, including an insight into the design and principle of operation of the coherence-controlled holographic microscope. Next a brief description of current cancer treatments introduces new class of anti-cancer drug candidates designated migrastatics that should impair cancer cell migration and thus prevent late metastases formation. The main part of the thesis deals with the design of the experimental observation procedure and data processing using the holographic incoherent Quantitative Phase Imaging technique. The last part of the thesis is the subsequent analysis of the migrastatic effect of the selected drugs on cancer cells in vitro and the comparison of the obtained results with existing studies. Finally, the objectives with magnification 4x and 10x were evaluated and compared for starting “the Rapid Assessment of Cell Growth and Migration”.
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Identification of changes in the behaviour of tumour cells in low concentrations of migrastatics by coherence-controlled holographic microscope
Zábranská, Magdaléna ; Netíková,, Irena Štenglová (referee) ; Veselý, Pavel (advisor)
The subject of the diploma thesis was using of Coherence-Controlled Holographic Microscope in the research of the effect of migrastatics at different concentrations on tumor cells and the interpretation of quantitative phase imaging data. The theoretical introduction is divided into two parts. The first part is devoted to CCHM, which briefly describes the history of holographic microscopy, the principle and a detailed description of the main parts of the microscope. The second part is an introduction to the issue of cancer treatment and the introduction of migrastatic drugs. The experimental part of the work is the analysis of the influence of migrastatics on two tumor lines (H1299 and HT1080) from the measured time-lapse records taken by a holographic microscope. The effect of drugs was evaluated using characteristics describing morphological and dynamic cell changes in the SophiQ software. The data were then graphically and statistically processed to compare and evaluate the effect of migrastatics at low levels. In conclusion of the work, the findings were summarized and selected characteristics were proposed to analyze the impact of migrastatics.
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Analysis of changes in the phenotype of tumour cells induced by migrastatics in quantitative phase imaging
Kolínková, Veronika ; Netíková,, Irena Štenglová (referee) ; Veselý, Pavel (advisor)
This thesis focuses on the observation of living cells using the non-invasive methods of quantitative phase imaging (hiQPI). The imaging is enabled by a coherence-controlled holographic microscope (CCHM) developed in the Laboratory of Experimental Biophotonics at the VUT. Using this imaging technology, morphological changes of A549 and MCF7 after application of potential migrastatic drugs tumor cells are evaluated in the experimental part of the thesis. Migrastatics are defined as already approved drugs that could prevent the migration of cancer cells from the primary tumor and thus prevent metastasis. The RAC-GM (Rapid Assessment of Cell Growth and Migration in Vitro) method was chosen to assess their effect on tumor cells.
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