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Thin metal films for detection and preconcentration
Podešva, Pavel ; Foret, František
We have tested CE separation of selected samples in capillaries with polydopamine modified surface. The capillaries were modified by polydopamine or polydopamine with an additive. The polydopamine coating with additive represents a simple and effective procedure for capillary alteration by another modificator such as hydroxyethyl cellulose, hydroxypropyl cellulose etc. In this work, we represent separation data from PrinCE system with UV detection for peptides, proteins and oligosaccharides labeled by 2-aminobenzoic acid.
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Polydopamine coated capillaries for CE separations
Partyka, Jan ; Foret, František ; Vu, T. T. ; Sudor, J.
We have tested CE separation of selected samples in capillaries with polydopamine modified surface. The capillaries were modified by polydopamine or polydopamine with an additive. The polydopamine coating with additive represents a simple and effective procedure for capillary alteration by another modificator such as hydroxyethyl cellulose, hydroxypropyl cellulose etc. In this work, we represent separation data from PrinCE system with UV detection for peptides, proteins and oligosaccharides labeled by 2-aminobenzoic acid.
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Multi-step synthesis of caspase-3 sensor based on Förster resonance energy transfer
Lišková, Marcela ; Klepárník, Karel ; Pazdera, P. ; Foret, František
Programmed cell death or apoptosis is regulated process of cell suicide. The central role in apoptosis play cysteine proteases called caspases. Caspases recognize tetra-peptide sequences Asp-Glu-Val-Asp (DEVD) on their substrates and hydrolyze peptide bonds after aspartic acid residues. Various techniques for the determination of caspase-3 are commercially available e.g. Enzyme Linked Immuno-Sorbent Assay (ELISA), Western blotting or flow cytometric analysis. The products of the cleavage can be detected by spectrophotometry, fluorimetry, chemiluminescence (CL) or ELISA. In this work, we suggested fluorescent sensor based on Förster Resonance Energy Transfer (FRET) to determine caspase-3 in cell nucleus or cytoplasm by apoptosis for very fast analysis by fluorescence microscopy without sample destroying.
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Sensor based on Förster resonance energy transfer with quantum dot
Lišková, Marcela ; Datinská, Vladimíra ; Klepárník, Karel ; Foret, František
Förster resonance energy transfers (FRET) are based on the nonradiative transfer of energy between a donor and acceptor dye in the distance 1-10 nm. With respect to this, quantum dots (QDs) with a high absorbance can serve as suitable donors of energy in FRET. Here, we present architectures of two sensors based on QD conjugates useful in imaging and diagnostics of cancer. One sensor, based on an oligonucleotide conjugate, can be used for the detection of complementary oligonucletide chains. The other was designed for the detection of enzymes inside cells.
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Photolithographic technique for controlled fabrication of metal structures
Jusková, Petra ; Hegrová, Jitka ; Foret, František
We are describing two strategies for controlled (geometry and dimension) preparation of metal micro-nano structures. Both techniques are based on photolithographic processes with selective pattern transfer. First technique utilizes agarose gel containing photosensitive silver chloride as the photosensitive layer. Silver structures are formed after irradiation of the gel through the photolithographic mask which defines shape of the resulting structures. Second approach utilizes photolithographic process to form particles from vacuum deposited thin metal layer deposited over soluble sacrificial layer.
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Iron oxide nanoparticle modified monolithic pipette tips for selective enrichment of phosphopeptides
Křenková, Jana ; Foret, František
We have developed iron oxide nanoparticle modified monolithic pipette tips for selective and efficient enrichment of phosphopeptides. Iron oxide nanoparticles were synthesized using a co-precipitation method and stabilized by citrate ions. A stable coating of nanoparticles was obtained via multivalent interactions of citrate ions on the nanoparticle surface with a quaternary amine functionalized surface of the methacrylate based monolithic tips. The performance of the developed and commercially available tips was demonstrated with the enrichment of phosphorylated peptides from tryptic digests of caseins followed by MALDI/MS detection.
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Thin metal films in resistivity-based chemical sensing
Podešva, Pavel ; Foret, František
Measurement of basic electric properties (current, voltage, resistance) is the most frequent technique in practically all scientific disciplines. Direct electric measurements are simple, precise and provide very large dynamic range. Since the related equipment is simple and inexpensive sensors based on electric measurements are very attractive for many applications in chemistry, environmental sciences or medicine. In this work, we present resistivity sensor for direct measurement of thiols in liquid samples.
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