|
|
Coherence Controlled Holographic Microscope with the digital optics
Vavřinová, Jana ; Jákl, Petr (referee) ; Dostál, Zbyněk (advisor)
The Digital Micromirror Device (DMD) technology has been developed especially for Digital Light Processing projectors, which allow the image projection. After this succesful implementation, and thanks to the commercial availibility and low initial cost of the DMD chip, a wide range of other applications became possible. Besides, it may be used in microscopy as a spatial light modulator. For example in Coherence-Controlled Holographic Microscope (CCHM) that finds its use especially for imaging and measurement of live-cell dynamic processes. The DMD chip placed in the illumination part of CCHM allows for broadening the application possibilities. Namely it could be different illumination mode experiments or tomographic applications. The master's thesis deals with the optical design of CCHM with digital optics, i. e. DMD chip. The selection of optical elements for CCHM, the experimental verification of the imaging setup and the process of designing the illumination part are described in detail. In the end, the analysis of different designs for illumination setup with the digital optics in object arm is carried out and the results are compared.
|
|
| |
|
|
Analysis of Microscopic Images of Cancer Cells
Vičar, Tomáš ; Matula,, Petr (referee) ; Sladoje, Natasa (referee) ; Kolář, Radim (advisor)
Tato disertační práce je zaměřena na analýzu různých forem mikroskopických obrazových dat nádorových buněk (statické 2D snímky, statické 3D obrazy, 2D časosběrné zobrazování živých buněk). Hlavní pozornost je věnována datům získaným koherencí řízeným holografickým mikroskopem, který je relativně novou modalitou schopnou kotrastních záznamů živých buněk bez barvení (label-free) a poskytuje kvantitativní informaci (kvantitativní fázové zobrazení - QPI). V práci je popsán základní postup analýzy těchto snímků a jsou vytvářeny nové metody a zdokonalovány metody pro jednotlivé kroky této analýzy. Největší část práce je věnována segmentaci buněk, kde jsou shrnuty klasické metody i metody založené na hlubokém učení. Jsou také vyvinuty nové metody vhodné právě pro QPI data. Část práce je také věnována segmentaci 3D fluorescenční jader a detekci DNA zlomů pomocí hlubokého učení. Práce se zabývá i dalším zpracování v podobě sledování buněk, extrakce příznaků a následné analýze, kde je detekována buněčná smrt a jsou vytvořeny vhodné interpretovatelné příznaky pro klasifikaci buněčné smrti na apoptickou a lytickou. Celkově tato práce přispívá k rozvoji jednotlivých kroků analýzy obrazu nádorových buněk a odráží současný pokrok v oblasti analýzy obrazu, zejména přístupy hlubokého učení, což je také demonstrováno na několika výzkumných aplikacích.
|
|
|
Testing of multimodal holographic microscope in reflection mode
Kachtík, Lukáš ; Slabý, Tomáš (referee) ; Křížová, Aneta (advisor)
The thesis is focused on testing of reflection mode of Multimodal holographic microscope constructed in cooperation with TESCAN company. The assessment of axial and lateral resolution, verification of axial intensity response and documentation errors and deficiencies found while measuring are included in the testing procedure. In the conclusion, the Multimodal holographic microscope is compared to an atomic force microscop. A proper understanding of microscope operation and a formulation of an adjustment procedure were essential components of this work.
|
|
|
Automated Procedures for Coherence Controlled Holographic Microscope
Dostál, Zbyněk ; Štarha, Pavel (referee) ; Jákl, Petr (referee) ; Chmelík, Radim (advisor)
Coherence-Controlled Holographic Microscope (CCHM) and a Fluorescence Holographic Microscope (FHM) were developed particularly for quantitative phase imaging and measurement of live cell dynamics, which used to be a subject of digital holographic microscopy (DHM). CCHM and FHM in low-coherence mode extend capabilities of DHM in the study of living cells. However, this advantage following from the use of low coherence is accompanied by increased sensitivity of the system to its correct alignment. Therefore, the introduction of an automatic self-correcting system is inevitable. Accordingly, in the thesis, the theory of a suitable control system is derived and the design of an automated alignment system for both microscopes is proposed and experimentally proved. The holographic signal was identified as a significant variable for guiding the alignment procedures. On this basis the original basic realignment algorithms were proposed, which encompasses the processes for initial and advanced alignment as well as for long-term maintenance of the microscope aligned state. Automated procedures were implemented in both microscopes unique set of robotic mechanisms designed and built within the frame of the thesis work. All of the procedures described in the thesis were in real experimentally proved at real microscopes in the experimental biophotonics laboratory. In addition, the control software, which contains the needed automated procedures, was developed for FHM.
|
|
|
Analysis of Microscopic Images of Cancer Cells
Vičar, Tomáš ; Matula,, Petr (referee) ; Sladoje, Natasa (referee) ; Kolář, Radim (advisor)
Tato disertační práce je zaměřena na analýzu různých forem mikroskopických obrazových dat nádorových buněk (statické 2D snímky, statické 3D obrazy, 2D časosběrné zobrazování živých buněk). Hlavní pozornost je věnována datům získaným koherencí řízeným holografickým mikroskopem, který je relativně novou modalitou schopnou kotrastních záznamů živých buněk bez barvení (label-free) a poskytuje kvantitativní informaci (kvantitativní fázové zobrazení - QPI). V práci je popsán základní postup analýzy těchto snímků a jsou vytvářeny nové metody a zdokonalovány metody pro jednotlivé kroky této analýzy. Největší část práce je věnována segmentaci buněk, kde jsou shrnuty klasické metody i metody založené na hlubokém učení. Jsou také vyvinuty nové metody vhodné právě pro QPI data. Část práce je také věnována segmentaci 3D fluorescenční jader a detekci DNA zlomů pomocí hlubokého učení. Práce se zabývá i dalším zpracování v podobě sledování buněk, extrakce příznaků a následné analýze, kde je detekována buněčná smrt a jsou vytvořeny vhodné interpretovatelné příznaky pro klasifikaci buněčné smrti na apoptickou a lytickou. Celkově tato práce přispívá k rozvoji jednotlivých kroků analýzy obrazu nádorových buněk a odráží současný pokrok v oblasti analýzy obrazu, zejména přístupy hlubokého učení, což je také demonstrováno na několika výzkumných aplikacích.
|
|
|
Coherence Controlled Holographic Microscope with the digital optics
Vavřinová, Jana ; Jákl, Petr (referee) ; Dostál, Zbyněk (advisor)
The Digital Micromirror Device (DMD) technology has been developed especially for Digital Light Processing projectors, which allow the image projection. After this succesful implementation, and thanks to the commercial availibility and low initial cost of the DMD chip, a wide range of other applications became possible. Besides, it may be used in microscopy as a spatial light modulator. For example in Coherence-Controlled Holographic Microscope (CCHM) that finds its use especially for imaging and measurement of live-cell dynamic processes. The DMD chip placed in the illumination part of CCHM allows for broadening the application possibilities. Namely it could be different illumination mode experiments or tomographic applications. The master's thesis deals with the optical design of CCHM with digital optics, i. e. DMD chip. The selection of optical elements for CCHM, the experimental verification of the imaging setup and the process of designing the illumination part are described in detail. In the end, the analysis of different designs for illumination setup with the digital optics in object arm is carried out and the results are compared.
|
|
|
Automated Procedures for Coherence Controlled Holographic Microscope
Dostál, Zbyněk ; Štarha, Pavel (referee) ; Jákl, Petr (referee) ; Chmelík, Radim (advisor)
Coherence-Controlled Holographic Microscope (CCHM) and a Fluorescence Holographic Microscope (FHM) were developed particularly for quantitative phase imaging and measurement of live cell dynamics, which used to be a subject of digital holographic microscopy (DHM). CCHM and FHM in low-coherence mode extend capabilities of DHM in the study of living cells. However, this advantage following from the use of low coherence is accompanied by increased sensitivity of the system to its correct alignment. Therefore, the introduction of an automatic self-correcting system is inevitable. Accordingly, in the thesis, the theory of a suitable control system is derived and the design of an automated alignment system for both microscopes is proposed and experimentally proved. The holographic signal was identified as a significant variable for guiding the alignment procedures. On this basis the original basic realignment algorithms were proposed, which encompasses the processes for initial and advanced alignment as well as for long-term maintenance of the microscope aligned state. Automated procedures were implemented in both microscopes unique set of robotic mechanisms designed and built within the frame of the thesis work. All of the procedures described in the thesis were in real experimentally proved at real microscopes in the experimental biophotonics laboratory. In addition, the control software, which contains the needed automated procedures, was developed for FHM.
|
|
|
Automated Procedures for Coherence Controlled Holographic Microscope
Dostál, Zbyněk ; Chmelík, Radim (advisor)
Coherence-Controlled Holographic Microscope (CCHM) and a Fluorescence Holographic Microscope (FHM) were developed particularly for quantitative phase imaging and measurement of live cell dynamics, which used to be a subject of digital holographic microscopy (DHM). CCHM and FHM in low-coherence mode extend capabilities of DHM in the study of living cells. However, this advantage following from the use of low coherence is accompanied by increased sensitivity of the system to its correct alignment. Therefore, the introduction of an automatic self-correcting system is inevitable. Accordingly, in the thesis, the theory of a suitable control system is derived and the design of an automated alignment system for both microscopes is proposed and experimentally proved. The holographic signal was identified as a significant variable for guiding the alignment procedures. On this basis the original basic realignment algorithms were proposed, which encompasses the processes for initial and advanced alignment as well as for long-term maintenance of the microscope aligned state. Automated procedures were implemented in both microscopes unique set of robotic mechanisms designed and built within the frame of the thesis work. All of the procedures described in the thesis were in real experimentally proved at real microscopes in the experimental biophotonics laboratory. In addition, the control software, which contains the needed automated procedures, was developed for FHM.
|
|
| |
guest ::
