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Holographic module for a light microscopy
Škrabalová, Denisa ; Slabý, Tomáš (referee) ; Dostál, Zbyněk (advisor)
The new arrangement of the off-axis holographic module, which is using polarizationactive diffraction grating divides signal into reference and subject wave of an interferometer based on their polarization. However, current design of the module does not have a possibility to tune a length of the optical paths. Thus the inability to tune optical paths leads to a reduced quality of interference structure during observation of biological samples. The current module is only suitable for technical applicating due to this limitation. Possibility of tuning branches is key step in biological applications. Therefore a new computer-controlled module is created in order to enable use for biological samples.
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Superresolution in holographic microscopy
Říha, René ; Komrska, Jiří (referee) ; Chmelík, Radim (advisor)
An approach that uses a two dimensional phase grating in certain distance behind a specimen to enhance the resolution in digital holographic microscopy is proposed. By theoretical considerations and with simulations the properties of new the arrangement are explored and the method of numerical reconstruction is described. The proposed approach is verified by electromagnetic simulation. Technical aspects and potential difficulties aren't involved.
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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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Fabrication and characterization of nanostructures with functional properties in the field of plasmonics
Babocký, Jiří ; Detz, Hermann (referee) ; Vala,, Milan (referee) ; Dub, Petr (advisor)
Tato dizertční práce se zabývá výrbou a charakterizací plasmonických nanostruktur. Její první část začíná krátkým úvodem do plasmoniky s navazujícím přehledem metod, které jsou v dnešní době nejčastěji používány k výrobě a charakterizaci plasmonických nanostruktur. Druhá část se pak zaměřuje na samotný výzkum, který byl v rámci PhD studia realizován. Cílem prvních experimentů bylo prozkouat možnosti použití elektronové litografie za variabilního tlaku v procesní komoře pro výrobu plasmonických nanostruktur na nevodivých substrátech jako je např. sklo. Jelikož se jedná o materiály, které jsou velice často používány k přípravě plasmonických struktur pacujících v oblasti viditelného světla. Druhá sekce pak diskutuje některé specifické aspekty přípravy plasmonických mikrostruktur elektronovou litografií pro THz oblast. Poslední část se pak zaměřuje na funkční vlastnosti plasmonických nanostruktur, převážně pak na kvantitativní charakterizaci fáze dalekého pole indukovaného plasmonickými nanostrukturami a jejich aplikacemi v oblasti optických metapovrchů - uměle připravených povrchů, které mohou být použity jako planární optické komponenty. Práce demonstruje a diskutuje různé experimentální přístupy použití mimoosové holografické mikroskopie pro jejich charakterizaci.
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Application of the FDTD technique to modelling of imaging in biophotonics
Říha, René ; Richter, Ivan (referee) ; Petráček, Jiří (advisor)
This thesis deals with the problem of practical application of FDTD technique for simulation of image formation in coherence controlled holographic microscope. Various ways for obtaining scattering matrices are explored in detail and the optimal technique based on a rigorous calculation of the far field is proposed. The scattering matrix, containing information about the observed sample, is subsequently used in analytic calculation of holographic signal; two levels of approximation of pupil function are also evaluated. The results are compared with a traditional approach based on Rytov approximation resulting in specification of the parameter domain where the approximation is applicable. Based on the simulations of the microscope, the dependence of axial resolution on apertures of the objective and the condenser and sensitivity of the signal to changes of refractive index of the sample is also studied.
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Piezo stage for CCHM microscope
Motyčka, Vít ; Cejpek, Zdeněk (referee) ; Dostál, Zbyněk (advisor)
For new applications of three-dimensional imaging using a coherence-controlled holographic microscope, it is necessary to design a stage for samples that allows precise movement of the sample in the axial axis in a range larger than those normally offered by the manufacturers of these stages. This fact is supported by a search of commercially available products. Based on the requirements, two conceptual designs of the stage are determined. A suitable concept is converted into a design solution and is equipped with suitable piezo actuators. Production documentation is created. The stage is manufactured and assembled. Its correct function is verified by testing.
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Homogenized illumination source for microscopy
Riznerová, Petra ; Slabý, Tomáš (referee) ; Dostál, Zbyněk (advisor)
This thesis deals with the design of an illumination system for homogeneous illumination of the sample of a coherence-controlled holographic microscope. The theoretical part of the thesis briefly describes the principle of interference and holographic microscopy in connection with the use of a suitable illumination system for microscopy, including methods of light homogenization. The definition of the parameters of the new miniaturized source is based on this part. The work includes two designs of an illumination system with different use of microlens arrays. Optical simulations are performed, which verify the correct selection of all optical elements of the illumination system. Following the optical models, optomechanical designs are created. These are compiled and tested. The light efficiency of the sources and the distribution of light intensity values in their output planes are measured.
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Control and automation of the autofocus optical unit for a transmission holographic microscope
Paták, Marek ; Bouchal, Petr (referee) ; Dostál, Zbyněk (advisor)
The bachelor's thesis deals with the automation of the autofocus unit for a transmission holographic microscope. The work describes the construction of the existing autofocusing unit, which is supplemented by the robotization of the collimator using a belt mechanism. The work presents an algorithm for automating the collimator. An algorithm for evaluating the microscope detuning is also described. A graphical interface is created for these functions. The algorithms were experimentally tested on a laboratory setup for further integration into the control program of the holographic microscope.
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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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Stabilization of the mutual position of the focal plane and the observed object in the holographic microscope
Ha Trinh, Hung Son ; Brada, Michal (referee) ; Dostál, Zbyněk (advisor)
Koherencí řízený holografický mikroskop (CCHM) umožňuje studium nativních biologických vzorků v reálném čase. Při dlouhodobých pozorováních ovšem dochází k rozostřování sledovaných objektů. Tento nežádoucí jev je pravděpodobně způsoben změnami vzájemné polohy předmětové roviny a pozorovaného předmětu. Předpokládá se, že za nestálostí vzájemných poloh stojí tepelné změny uvnitř CCHM. Vznikla proto potřeba vyvinout hardwarový autofokusační systém, který by doplňoval v současnosti implementovaný softwarový autofokus a mohl by vykompenzovat jeho nedostatky. V práci je nejprve stručně popsána optika CCHM. Poté je vysvětlen princip aktuálního řešení doostřování. Posléze je představeno několik metod detekce a monitorování posunutí, kterých lze využít ve zpětnovazebné smyčce k udržování definované vzájemné polohy. Po zvážení výhod a nevýhod je pro praktickou část práce vybrána právě jedna metoda, na základě které je navrženo, sestrojeno a experimentálně otestováno zařízení zajišťující stabilizaci vzájemné polohy předmětové roviny a pozorovaného předmětu.
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