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Master's Thesis
Author of thesis: Ing. Jana Čolláková
Acad. year: 2010/2011
Supervisor: Ing. Martin Antoš, Ph.D.
Reviewer: Ing. Jan Ježek, Ph.D.
The perfusion chamber for long term observing of live cells by the means the Coherence-Controlled Holographic Microscope (CCHM) was designed. CCHM was built and designed at the Laboratory of the optical microscopy at the Institute of Physical Engineering, Brno University of Technology. CCHM can quantitatively evaluate dynamical changes inside live cells thanks to the quantitative information about phase shift in each pixel of the image. In order to demonstrate advantages of CCHM experimentally, it is important to keep the live cells in the good conditions. This is made by adding the fresh cultivation medium for studied cells directly in the microscope. In contrast to the stationary chamber the perfusion chamber allows both the cultivation medium exchange and the application of biological reagents without the necessity of removing the chamber from the microscope. Therefore we can study the vital signs of cells before and after the application of reagents. An original perfusion system with accessories compatible with CCHM was designed. The design is based on the previously published perfusion system solutions that are referred to in this thesis. The flow characteristics and medium exchange process was discussed and a modification of the internal geometry, based on numerical simulations, was introduced. The applicability of this perfusion chamber has been proven for the CCHM and even for different types of microscopes. The reactions of tumor and epithelial cells during the change of the environment from the cultivation medium to the physiologically solution were studied.
Coherence-controlled holographic microscope, live cell imaging, perfusion chamber.
Date of defence
20.06.2011
Result of the defence
Defended (thesis was successfully defended)
Grading
A
Language of thesis
Czech
Faculty
Fakulta strojního inženýrství
Department
Institute of Physical Engineering
Study programme
Applied Sciences in Engineering (N3901-2)
Field of study
Precise Mechanics and Optics (M-PMO)
Composition of Committee
prof. RNDr. Tomáš Šikola, CSc. (předseda) prof. RNDr. Miroslav Liška, DrSc. (místopředseda) prof. RNDr. Bohumila Lencová, CSc. (člen) doc. RNDr. Josef Kuběna, CSc. (člen) prof. RNDr. Jiří Komrska, CSc. (člen) prof. RNDr. Pavel Zemánek, Ph.D. (člen) prof. RNDr. Petr Dub, CSc. (člen) prof. RNDr. Radim Chmelík, Ph.D. (člen) prof. Ing. Ivan Křupka, Ph.D. (člen) prof. RNDr. Jiří Spousta, Ph.D. (člen) RNDr. Antonín Fejfar, CSc. (člen)
Supervisor’s reportIng. Martin Antoš, Ph.D.
Grade proposed by supervisor: B
Reviewer’s reportIng. Jan Ježek, Ph.D.
Grade proposed by reviewer: B
Responsibility: Mgr. et Mgr. Hana Odstrčilová