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Deconvolution fluorescence microscopy of yeast cells
Štec, Tomáš ; Plášek, Jaromír (advisor) ; Heřman, Petr (referee)
Title: Deconvolution fluorescence microscopy of yeast cells Author: Tomáš Štec Department: Institute of Physics of Charles University Supervisor: prof. RNDr. Jarmoír Plášek, CSc., Institute of Physics of Charles Uni- versity Abstract: Fluorescence microscopy presents an fast and cheap alternative to more advanced imaging methods like confocal and electron microscopy, even though it is subject to heavy image distortion. It is possible to recover most of the original distortion-free image using deconvolution in computer image processing. This al- lows reconstruction of 3D structure of studied objects. Deconvolution procedure of NIS Elements AR program undergoes an thorough inspection in this diploma the- sis. It is then applied on restoration of 3D structure of calcofluor stained cell wall of budding yeast Saccharomyces cerevisiae. Changes of the structure of the cell wall during cell ageing are being examined. Cell wall of aged cells shows increased surface roughness and even ruptures at the end of cell life. Keywords: fluorescence, microscopy, deconvolution, NIS Elements AR, calcofluor, yeast, cell wall, ageing
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Deconvolution fluorescence microscopy of yeast cells
Štec, Tomáš ; Plášek, Jaromír (advisor) ; Heřman, Petr (referee)
Title: Deconvolution fluorescence microscopy of yeast cells Author: Tomáš Štec Department: Institute of Physics of Charles University Supervisor: prof. RNDr. Jarmoír Plášek, CSc., Institute of Physics of Charles Uni- versity Abstract: Fluorescence microscopy presents an fast and cheap alternative to more advanced imaging methods like confocal and electron microscopy, even though it is subject to heavy image distortion. It is possible to recover most of the original distortion-free image using deconvolution in computer image processing. This al- lows reconstruction of 3D structure of studied objects. Deconvolution procedure of NIS Elements AR program undergoes an thorough inspection in this diploma the- sis. It is then applied on restoration of 3D structure of calcofluor stained cell wall of budding yeast Saccharomyces cerevisiae. Changes of the structure of the cell wall during cell ageing are being examined. Cell wall of aged cells shows increased surface roughness and even ruptures at the end of cell life. Keywords: fluorescence, microscopy, deconvolution, NIS Elements AR, calcofluor, yeast, cell wall, ageing
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Fluorescence spectroscopy of calcofluor stained yeast cell suspensions.
Dostál, Marek ; Plášek, Jaromír (advisor) ; Heřman, Petr (referee)
Title: Fluorescence spectroscopy of calcofluor stained yeast cell suspensions Author: Marek Dostál Department: Fyzikální ústav UK Supervisor: prof. RNDr. Jaromír Plášek CSc. Abstract: We were finding and we found fast and easy fluorescence spectroscopy method which we can use for cell wall research of different yeast types that were cultivated on various medium. Gauging was under way in solution of different ionic forces. The growth medium has the greatest effect on quality of cell wall. We use calcofluor as fluorescent colour. After exposing calcofluor by UV (λex ≈ 365 nm) calcofluor emites blue fluorescence with maximum intensity around 420 nm in dependence on suspension concentration. We must be careful about inner filter that can influence our result during research. We validated diversity of cell wall structures in various yeast types by ratio spectral method (spectral fingerprint). Keywords: Fluorescence, spectroscopy, cell wall, calcofluor, inner filter.
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