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Preparation of luminescent nanoparticles for 3D imaging
Smolka, Rastislav ; Hrabal, Michal (referee) ; Vala, Martin (advisor)
The aim of this bachelor thesis was to study a suitable methodology for the preparation of fluminiscent nanoparticles and to characterize their optical properties associated with various physical properties of environment. Also, their potential use as a means of monitoring biological preparations in 3D microscopy using multiphoton absorption. In particular, we deal with nanoparticles prepared from N,N-diphenylamino-2,5-diphenyl-1,4-distyrylbenzene cyanide and their fluorescence and absorption spectra. Using the dynamic light scattering method and fluorescence spectroscopy, the dependence of the size of the prepared nanoparticles, the change in their fluorescence properties and the change in the quantum yield is observed, depending on the changing environmental properties. Data from multifoton absorption measurements demonstrate that this substance responds to multifoton excitation. It has been shown that this substance retains its unique fluorescence properties also in the form of nanoparticles and thus appears to be a suitable candidate for the observation of biological preparations using multifoton fluorescence microscopy. Therefore, the continuation of this work could be the mere determination of the total multifoton absorption cross section of these prepared nanoparticles.
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Manufacturing of micro-structures by two-photon polymerization
Skalický, Jiří ; Petrůj, Jaroslav (referee) ; Richtera, Lukáš (advisor)
Utilization of focused laser beam for photo-polymerization represents an elegant and fast method how to manufacture microstructures in dimensions from hundreds of nanometres to tens of micrometres. Two-photon photo-polymerization is used with advantage to manufacture microstructures with sub-micrometre details. In this case the laser wavelength is about two times longer comparing to the spectral region of the highest photoresist sensitivity and the two-photon photo-polymerization is initiated only in the positions of sufficient optical intensity. Precise and controlled positioning of the photoresist with respect to the laser focus ensures only illumination of areas of future microstructure and initialization of photo-polymerization here. Fine details of manufactured microstructures depends, except the laser beam focusing, also on the chosen photoresist and quenching of polymerization in non-illuminated areas.
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Study of optical properties of organic materials for nonlinear optical applications
Hrbková, Silvie ; Hrabal, Michal (referee) ; Vala, Martin (advisor)
This work deals with a development of a method for determination of two-photon parametres and its application. The theoretical part includes a description of two-photon absorption processes, most frequently used methods to determine two-photon cross sections and a summary of molecules researched to date in the field. The molecules studied in this work are Rhodamin B and Rhodamin 6G in ethanol, which are applied as standards for the implementation of the method. After that, Diketopyrrolopyrroles U24, U37, U49 and U50 dissolved in DMSO, toluene, acetone and chloroform, are researched. Basic optical characteristics - absoption and emission spectra were experimentally acquired for all the materials. Two-photon characteristics were obtained using a picosecond laser and a Two-Photon Excited Fluorescence (TPEF) method. TPA cross-sections and TPA spectra of Rhodamin B and Rhodamin 6G were measured and discussed. Subsequently, this work focuses on absorption and emission solvatation processes of the selected Diketopyrrolopyrroles. These processes indicate their two-photon efficiency. Finally, TPA spectra of the substances are discussed. Contribution of this work is a developed method for measuring the two-photon characteristics suitable for further use. The outcomes achived, enable extensive research of the Diketopyrrolopyrrols characteristics with the use of the measured standards of the Rhodamin B and Rhodamin 6G.
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Two-photon photopolymerization with multiple laser beams
Skalický, Jiří ; Pilát,, Zdeněk (referee) ; Jákl,, Petr (advisor)
Photopolymerization is a technique used to create surface structures or microobjects from a photoresist. This process is started by illuminating the sample with a light of proper wavelength absorbed by the resist. After exposure, the sample is processed according to the type of the photoresist – be it heating, treating with developer or just washing the unaffected monomer with some reagent. Focused femtosecond laser beam with double wavelength can be used in the process. Short pulse length with high photon density starts two-photon absorption localized in the vicinity of focal point. The method resolution is thus increased and details with 1/10 micrometer size can be created. Moreover, very short laser pulse decreases the heat affected zone and the risk of thermal initiation is minimized. Manufacturing of larger structures composed of tiny details with two-photon photopolymerization is time-demanding process. Therefore, we have complemented the optical setup with spatial light modulator (SLM), which splits the incoming laser beam into several beams with holograms dynamically generated by a computer. Polymerization can be thus performed by multiple foci simultaneously which can be used to create separated microparticles or periodical surface structures. Additional speed improvement of the process can be substitution of static configuration, requiring sample replacement after each exposition, with continuous setup using microfluidic channel steadily supplied with photoresist transported to the active region of the sample.
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Luminescent nanoparticles for 3D imaging
Smolka, Rastislav ; Mravec, Filip (referee) ; Vala, Martin (advisor)
The aim of this diploma thesis is to study the optical properties of new -conjugated molecules based on 1,4-di(4'-N, N-diphenylaminostyryl)benzene and their potential application in advanced imaging techniques of biological specimens, the so-called multiphoton microscopy. The thesis focuses mainly on the characterization of their optical properties and the determination of their two-photon absorption cross-section using a unique laser equipment. Furthermore, a suitable methodology for the preparation of nanoparticles from these molecules, their characterization and stability are also developed. The thesis also investigates the influence of structure on the optical properties of these molecules. The relation between the length of the conjugated system and the presence of substituents on the backbone has been shown for the optical properties of the molecules in the solvent, the position and shape of the two-photon absorption spectrum and the value of two-photon absorption cross-section. It has been shown that this substance retains its unique fluorescent properties even in the form of nanoparticles and therefore appears to be a suitable candidate for the observation of biological specimens using multiphoton fluorescence microscopy. The work contributes to the knowledge base for the design of the chemical structure of molecules with desired properties.
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Luminescent nanoparticles for 3D imaging
Smolka, Rastislav ; Mravec, Filip (referee) ; Vala, Martin (advisor)
The aim of this diploma thesis is to study the optical properties of new -conjugated molecules based on 1,4-di(4'-N, N-diphenylaminostyryl)benzene and their potential application in advanced imaging techniques of biological specimens, the so-called multiphoton microscopy. The thesis focuses mainly on the characterization of their optical properties and the determination of their two-photon absorption cross-section using a unique laser equipment. Furthermore, a suitable methodology for the preparation of nanoparticles from these molecules, their characterization and stability are also developed. The thesis also investigates the influence of structure on the optical properties of these molecules. The relation between the length of the conjugated system and the presence of substituents on the backbone has been shown for the optical properties of the molecules in the solvent, the position and shape of the two-photon absorption spectrum and the value of two-photon absorption cross-section. It has been shown that this substance retains its unique fluorescent properties even in the form of nanoparticles and therefore appears to be a suitable candidate for the observation of biological specimens using multiphoton fluorescence microscopy. The work contributes to the knowledge base for the design of the chemical structure of molecules with desired properties.
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|
|
Preparation of luminescent nanoparticles for 3D imaging
Smolka, Rastislav ; Hrabal, Michal (referee) ; Vala, Martin (advisor)
The aim of this bachelor thesis was to study a suitable methodology for the preparation of fluminiscent nanoparticles and to characterize their optical properties associated with various physical properties of environment. Also, their potential use as a means of monitoring biological preparations in 3D microscopy using multiphoton absorption. In particular, we deal with nanoparticles prepared from N,N-diphenylamino-2,5-diphenyl-1,4-distyrylbenzene cyanide and their fluorescence and absorption spectra. Using the dynamic light scattering method and fluorescence spectroscopy, the dependence of the size of the prepared nanoparticles, the change in their fluorescence properties and the change in the quantum yield is observed, depending on the changing environmental properties. Data from multifoton absorption measurements demonstrate that this substance responds to multifoton excitation. It has been shown that this substance retains its unique fluorescence properties also in the form of nanoparticles and thus appears to be a suitable candidate for the observation of biological preparations using multifoton fluorescence microscopy. Therefore, the continuation of this work could be the mere determination of the total multifoton absorption cross section of these prepared nanoparticles.
|
|
|
Two-photon photopolymerization with multiple laser beams
Skalický, Jiří ; Pilát,, Zdeněk (referee) ; Jákl,, Petr (advisor)
Photopolymerization is a technique used to create surface structures or microobjects from a photoresist. This process is started by illuminating the sample with a light of proper wavelength absorbed by the resist. After exposure, the sample is processed according to the type of the photoresist – be it heating, treating with developer or just washing the unaffected monomer with some reagent. Focused femtosecond laser beam with double wavelength can be used in the process. Short pulse length with high photon density starts two-photon absorption localized in the vicinity of focal point. The method resolution is thus increased and details with 1/10 micrometer size can be created. Moreover, very short laser pulse decreases the heat affected zone and the risk of thermal initiation is minimized. Manufacturing of larger structures composed of tiny details with two-photon photopolymerization is time-demanding process. Therefore, we have complemented the optical setup with spatial light modulator (SLM), which splits the incoming laser beam into several beams with holograms dynamically generated by a computer. Polymerization can be thus performed by multiple foci simultaneously which can be used to create separated microparticles or periodical surface structures. Additional speed improvement of the process can be substitution of static configuration, requiring sample replacement after each exposition, with continuous setup using microfluidic channel steadily supplied with photoresist transported to the active region of the sample.
|
|
|
Study of optical properties of organic materials for nonlinear optical applications
Hrbková, Silvie ; Hrabal, Michal (referee) ; Vala, Martin (advisor)
This work deals with a development of a method for determination of two-photon parametres and its application. The theoretical part includes a description of two-photon absorption processes, most frequently used methods to determine two-photon cross sections and a summary of molecules researched to date in the field. The molecules studied in this work are Rhodamin B and Rhodamin 6G in ethanol, which are applied as standards for the implementation of the method. After that, Diketopyrrolopyrroles U24, U37, U49 and U50 dissolved in DMSO, toluene, acetone and chloroform, are researched. Basic optical characteristics - absoption and emission spectra were experimentally acquired for all the materials. Two-photon characteristics were obtained using a picosecond laser and a Two-Photon Excited Fluorescence (TPEF) method. TPA cross-sections and TPA spectra of Rhodamin B and Rhodamin 6G were measured and discussed. Subsequently, this work focuses on absorption and emission solvatation processes of the selected Diketopyrrolopyrroles. These processes indicate their two-photon efficiency. Finally, TPA spectra of the substances are discussed. Contribution of this work is a developed method for measuring the two-photon characteristics suitable for further use. The outcomes achived, enable extensive research of the Diketopyrrolopyrrols characteristics with the use of the measured standards of the Rhodamin B and Rhodamin 6G.
|
|
|
Manufacturing of micro-structures by two-photon polymerization
Skalický, Jiří ; Petrůj, Jaroslav (referee) ; Richtera, Lukáš (advisor)
Utilization of focused laser beam for photo-polymerization represents an elegant and fast method how to manufacture microstructures in dimensions from hundreds of nanometres to tens of micrometres. Two-photon photo-polymerization is used with advantage to manufacture microstructures with sub-micrometre details. In this case the laser wavelength is about two times longer comparing to the spectral region of the highest photoresist sensitivity and the two-photon photo-polymerization is initiated only in the positions of sufficient optical intensity. Precise and controlled positioning of the photoresist with respect to the laser focus ensures only illumination of areas of future microstructure and initialization of photo-polymerization here. Fine details of manufactured microstructures depends, except the laser beam focusing, also on the chosen photoresist and quenching of polymerization in non-illuminated areas.
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