Miniaturization of devices to study chemical interactions and processes in liquid samples has led to the emergence of microfluidics and construction of lab-on-a-chip systems. Present work was devoted to implementation, development and testing of microfluidic systems with detection by confocal Raman microscopy and surface enhanced Raman scattering under the conditions of training department. Several options of performing standard macroscopic measurements in microscopic scales were explored. A method for measuring thermal stability of biopolymers in microsystems with contactless detection of temperature has been designed and tested. Furthermore, possibilites for studying the SERS effect within microfluidic channels were explored. It was demonstrated that the microfluidic chips provide promising opportunity to study hydrodynamics of liquids at microscopic level and chemical reactions and kinetics.

This work deals with fabrication and test of microfluidic chip for separating substances. For separation of substances is used electrophoresis and detection is by integrated electrodes.

The diploma thesis deals with cellular membrane model preparation on microfluidic devices. It summarizes means of microfluidic device fabrication, phospholipid bilayer formation mechanisms, optimization techniques and characterization methods of those systems. It focuses on free-standing planar lipid bilayers which are easily accessible by a number of different characterization methods and at the same time exhibit good stability and variability. The aim of this work is to design and prepare a microfluidic chip on which a planar lipid bilayer can be prepared. It therefore presents microfluidic device prepared by soft lithography of PDMS adapted for model membrane formation by self-assembly of phospholipids at the interface of aqueous and organic phases created by the architecture of the microfluidic device. Formation of the model membrane was visualized by optical microscopy and fluorescence-lifetime imaging microscopy.

Liposomes are versatile biocompatible and biodegradable carriers for a variety of medical applications. As the first nanoparticles, they have been approved for pharmaceutical use so far, and many liposome-based preparations are in clinical trials. Classical methods of liposome preparation represent potential limitations in technology transfer from laboratory to industrial scale. New, microfluidic techniques overcome these limitations and offer new possibilities for controlled, continuous preparation of liposomal particles in a laboratory and industrial scale. An important element in the development of new nanoparticle systems is their complex characterization and purification. In addition to the established chromatographic techniques, the Field flow fractionation technique, in particular the Asymmetrical flow Field-flow fractionation, is described. This relatively new technique in conjunction with the MALS/DLS/DAD-UV/dRI online detectors enables the purification and characterization of complex samples. The main advantage of this technique lies in the possibility of separation under native conditions, which plays an important role in the separation of biopolymers in particular. Separation in the “empty” channel then eliminates sample degradation due to unwanted interactions at the stationary phase-sample interface. The theoretical part of this thesis describes the possibilities of preparation, modification, and characterization of liposomal nanoparticles. For this purpose, optical methods based on dynamic light scattering, multi-angle dynamic light scattering and nanoparticle tracking analysis techniques are described, as well as a non-optical method using "particle by the particle" analysis, tunable resistive pulse sensing method. A separate chapter of the theoretical part is dedicated to the technique Asymmetrical flow Field-flow fractionation in connection with the above-mentioned detectors. Important results associated with this work are summarized in the attached scientific paper, together with the result summaries and the author's contributions.

In this master thesis I have been dealt with the design and construction of an instrumental platform that used positioning focused laser beam (so-called optical tweezers) for manipulation with living cells without their damage.

This work is focused on the development of a user friendly software interface using the LabViewTM environment that simplifies running of various experiments using laser micromanipulations and laser microspectroscopy of living microorganisms. Both techniques have been developing very fast for the last decade and belong to the growing group of contact-less and nondestructive techniques for manipulation and diagnostics of individual living microorganisms, cells, or viruses. Within this project we mastered the driving of peripheries, calibration of CCD scene, real-time image processing of the CCD scene, automatic selection of the cell for further laser processing, acquisition and processing of the Raman spectrum from living microorganisms. The final goal of our activity is fully automatic laser-based sorter of living cells depending on their chemical compositions. This work has been elaborated at the Institute of Scientific Instruments of the ASCR, v.v.i. under the supervision of prof. Pavel Zemanek.

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.