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Parallel single-cell analysis of active caspase-3/7 in apoptotic and non-apoptotic cells
Ledvina, Vojtěch ; Klepárník, Karel
Caspases are proteases that play key role in the process of apoptosis, the programmed\ncell death. Among them, caspase-3 and -7 are main executioner caspases that cleave\nmany vital proteins during apoptosis and after their widespread activation, the process\ncannot be reversed. To analyze caspase-3/7 activation within single cells, a miniaturized\ndevice for parallel analysis of eight samples was developed. The assay is based on the\nmodified luciferin-firefly luciferase bioluminescence (BL) system. Individual\nsuspended cells were collected and transferred into detection microvials using a\nmicromanipulator. The bioluminescence was detected using a photon counting head\nwith cooled photcathode. The LOD suitable for detection of active caspase-3/7 in both\napoptotic and non-apoptotic cells was reached.
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Development of microfluidic device for droplet generation and microparticle encapsulation
Křivánková, Jana ; Foret, František
Microfluidic devices combined with various bioanalytical and optical sensors proved\ntheir potential in chemical, biological, and biomedicine applications. At a droplet\nmicrofluidic device a fluid is divided into numerous droplets surrounded by a\ncontinuous immiscible fluid phase. Generated droplets serve then as\nvessels/microcapsules for delivering drugs, nutrients as well as for encapsulating of\nbiologically active particles and cells.\nDescribed experimental study deals with the development of a practical dropletbased\nmicrofluidic chip useful for production of monodisperse water/oil emulsion and\nfor encapsulation of thin-metal magnetic microsheets.
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CE-nESI/MS in electrode-free design: narrowing the separation channel to 5 μm
Týčová, Anna ; Foret, František
Capillary electrophoresis-nanospray/mass spectrometry (CE-nESI/MS) in electrodefree\narrangement represents a very simple instrumentation for analysis of biomolecules.\nSo far, CE-nESI/MS analyses in this design were limited to 10-75 μm ID of the\nseparation channel. Although the work with narrower dimensions brings several\nchallenges, there is a great potential for better sensitivity, improved separation power\nand lower sample consumption. This work is devoted to some practical aspects of\nexperiments in narrow bore channels and systematic evaluation of CE-nESI/MS\nanalyses conducted in capillaries of 25, 15 and 5 μm ID.
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Characterization of FRET sensor
Datinská, Vladimíra ; Klepárník, Karel ; Belšánová, B. ; Minárik, M. ; Foret, František
In this study, we present characterization of sensor based on Fӧrster resonance energy\ntransfer (FRET). The sensor is composed of ssDNA chain attached to a laboratory\nsynthesized quantum dot (QD). A complementary chain of a sample is labeled by a\nluminescent dye. When the dsDNA hybrid is formed, the energy from the QD (donor)\nis transferred to the dye (acceptor) and FRET is observed as a decrease of QD\nluminescence emission intensity and an increase of dye luminescence emission\nintensity.
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