Chemical vapor generation of cadmium: detector comparison
Sagapova, Linda; Štádlerová, Barbora; Svoboda, Milan; Kratzer, Jan; Musil, Stanislav
2021 - English
A chemical vapor generation system was connected to an in-house assembled atomic fluorescence spectrometer (AFS) equipped with a miniature diffusion flame atomizer in order to assess its potential to Cd determination. Subsequently, analytical performance of three spectrometric detectors was compared (AAS, AFS, ICP-MS) with respect to Cd determination after volatile species generation.
Keywords:
chemical vapor generation; cadmium; atomic fluorescence spectrometry
Available in digital repository of the ASCR
Chemical vapor generation of cadmium: detector comparison
A chemical vapor generation system was connected to an in-house assembled atomic fluorescence spectrometer (AFS) equipped with a miniature diffusion flame atomizer in order to assess its potential to ...
Bioluminescence determination of caspase-3/7 in single osteosarcoma cells
Killinger, Michael; Matalová, Eva; Klepárník, Karel
2021 - English
The protein heterogeneity at the single-cell level has been recognized to be vital for an understanding of various life processes during animal development. In addition, the knowledge of accurate quantity of relevant proteins at cellular level is essential for appropriate interpretation of diagnostic and therapeutic results. Here, we determined the level of active caspase-3/7 in physiological and pathological (cancer) conditions by our miniaturized instrumentation.
Keywords:
single-cell analysis; osteoblast; osteosarcoma
Available in digital repository of the ASCR
Bioluminescence determination of caspase-3/7 in single osteosarcoma cells
The protein heterogeneity at the single-cell level has been recognized to be vital for an understanding of various life processes during animal development. In addition, the knowledge of accurate ...
Epitachophoretic preconcentration of proteins
Hrušková, Helena; Voráčová, Ivona; Foret, František
2021 - English
In this work, we present a developed method for preconcentration of standards of proteins cytochrome c, myoglobin, and hemoglobin. Firstly, the electrolyte system was selected and optimized to provide low analysis time, narrow shape of the protein zone, and eliminate overheating at the center of the device. In the next step, the developed method was tested on the preconcentration of proteins: cytochrome c, myoglobin, and hemoglobin. After several adjustments myoglobin and hemoglobin were also uccessfully preconcentrated. After evaluation of pilot results, this technique provides up to fiftyfold preconcentration and recovery of over 70 % for each protein. The next plan is to preconcentrate proteins from complex biological matrices such as urine or blood plasma.
Keywords:
epitachophoresis; isotachophoresis; preparative concentration
Available on request at various institutes of the ASCR
Epitachophoretic preconcentration of proteins
In this work, we present a developed method for preconcentration of standards of proteins cytochrome c, myoglobin, and hemoglobin. Firstly, the electrolyte system was selected and optimized to provide ...
Separace VRA/RA ze vzorků pomocí isoelektrické fokusace v čipovém formátu (chip-IEF) pro potřeby kontroly zákazu biologických zbraní
Kubíček, O.; Duša, Filip; Šalplachta, Jiří; Lunerová, K.; Rosenbergová, K.
2021 - English
Metodika popisuje způsob separace bakteriálních vysoce rizikových a rizikových agens (VRA/RA) ze vzorků pomocí isoelektrické fokusace v čipovém formátu (chip-IEF). Metoda chip-IEF využívá nově vyvinutý unikátní prototyp zařízení pro vertikální isoelektrickou fokusaci v rozbíhavém toku v čipovém provedení, která na rozdíl od dosavadních postupů umožňuje i separaci bakterií. Separace pomocí chip-IEF funguje i pro šetrně inaktivované bakterie, což významně snižuje bezpečnostní rizika při manipulaci se vzorkem potenciálně obsahujícím VRA/RA.
Keywords:
isoelectric focusing; B-agens; chip
Fulltext is available at external website.
Separace VRA/RA ze vzorků pomocí isoelektrické fokusace v čipovém formátu (chip-IEF) pro potřeby kontroly zákazu biologických zbraní
Metodika popisuje způsob separace bakteriálních vysoce rizikových a rizikových agens (VRA/RA) ze vzorků pomocí isoelektrické fokusace v čipovém formátu (chip-IEF). Metoda chip-IEF využívá nově ...
Quantum dot luminescent probe for Caspase 3/7 imaging inside cells
Procházková, Markéta; Klepárník, Karel
2021 - English
Two step synthesis of luminescent probe based on ligand-exchange in the first step and the specific reaction of amino group in the second step was optimized. The luminescence properties of the final product were checked by time course of\nactive recombinant caspase protein reaction under the model conditions in fluorimeter. The novel luminescent probe enables long-time imaging of active caspases in living cells or tissues.
Keywords:
caspase; fluorescence; quantum dot
Available on request at various institutes of the ASCR
Quantum dot luminescent probe for Caspase 3/7 imaging inside cells
Two step synthesis of luminescent probe based on ligand-exchange in the first step and the specific reaction of amino group in the second step was optimized. The luminescence properties of the final ...
Detekce a identifikace aflatoxinů pomocí přenosného kapalinového chromatografu pro potřeby kontroly zákazu biologických zbraní
Šesták, Jozef; Lunerová, K.
2020 - Czech
Metodika popisuje způsob screeningové detekce a identifikace vybraných mykotoxinů aflatoxinů (aflatoxinů B1, B2, G1 a G2), ochratoxinů A, B a zearalenonu pomocí přenosného miniaturizovaného kapalinového chromatografu (miniLC) s UV/VIS a fluorescenčním detektorem (FLD). Cílem metodiky je poskytnout rychlou metodu pro zjištění/vyloučení přítomnosti uvedených mykotoxinů v neznámém vzorku. V případě pozitivního zjištění je nutné u daného vzorku provést potvrzení identity látek pomocí dalších instrumentálních metod (např. LC-MS). Metodika je určena zejména pro potřeby technické podpory SÚJB při kontrolní a dozorové činnosti v oblasti dodržování Zákona č. 281/1997 Sb. Method describes screening detection and identification of selected mycotoxins aflatoxins (aflatoxins B1, B2, G1 and G2), ochratoxins A, B and zearalenon by means of portable miniaturzed capillary liquid chromatograph (miniLC) with UV/VIS and fluorescence detector (FLD). The aim of the method is to provide rapid screening method for finding/excluding the presence of given mycotoxins in unknown samples. In case of positive finding, it is necessary to perform confirmation analysis using other instrumental methods (e. g. LC-MS). The method is intended namely for the needs of technical support of the State Office for Nuclear Safety for control and supervising activities in field of execution of the Act No. 281/1997 Coll (Biological Act).
Keywords:
miniaturized capillary chromatograph; aflatoxins; portable liquid chromatograph
Available on request at various institutes of the ASCR
Detekce a identifikace aflatoxinů pomocí přenosného kapalinového chromatografu pro potřeby kontroly zákazu biologických zbraní
Metodika popisuje způsob screeningové detekce a identifikace vybraných mykotoxinů aflatoxinů (aflatoxinů B1, B2, G1 a G2), ochratoxinů A, B a zearalenonu pomocí přenosného miniaturizovaného ...
Photochemical vapor generation of cobalt for detection by inductively coupled plasma mass spectrometry
Vyhnanovský, Jaromír; Musil, Stanislav
2020 - English
Volatile species of cobalt were generated in a flow-injection system, employing a high-efficiency flow-through UV photoreactor and a formic acid based medium. Optimal generation conditions were found as 10% (v/v) formic acid and 4 mol/L ammonium formate at a flow rate of 4 ml/min.
Keywords:
photochemical vapor generation; cobalt; inductively coupled plasma mass spectrometry
Available in digital repository of the ASCR
Photochemical vapor generation of cobalt for detection by inductively coupled plasma mass spectrometry
Volatile species of cobalt were generated in a flow-injection system, employing a high-efficiency flow-through UV photoreactor and a formic acid based medium. Optimal generation conditions were found ...
Photon-upconversion nanoparticles for single-molecule immunosensing of cancer biomarkers and bacteria
Farka, Z.; Mickert, M. J.; Hlaváček, Antonín; Poláchová, V.; Kostiv, Uliana; Pastucha, M.; Horák, Daniel; Gorris, H. H.; Skládal, P.
2020 - English
The recent progress in the field of immunoassays has been driven by introduction of various kinds of nanomaterials. In particular, photon-upconversion nanoparticles (UCNPs) proved to be excellent immunoassay labels due to their ability to emit light of shorter wavelengths under near-infrared excitation (anti-Stokes emission), which prevents autofluorescence, minimizes light scattering, and thus reduces the optical background interference. These unique photoluminescent properties allow counting of individual biomolecules labeled with UCNPs by conventional wide-field epiluminescence microscopy and enable the development of single-molecule (digital) immunoassays. We have introduced a novel label based on UCNPs conjugated with streptavidin via poly(ethylene glycol) and applied it in a digital upconversion-linked immunosorbent assay (ULISA) for the detection of a cancer biomarker prostate specific antigen (PSA). The digital readout based on counting of individual immunocomplexes improved the sensitivity 16× compared to conventional analog readout and allowed to reach a limit of detection (LOD) of 23 fg·mL−1 (800 aM). Human serum samples were successfully analyzed achieving an excellent correlation with electrochemiluminescence reference method. The conjugates of UCNPs with streptavidin are also suitable for the detection of pathogenic bacterium Melissococcus plutonius, the causative agent of honeybee disease European foulbrood. The ULISA assay provided an LOD of 340 CFU·mL−1 and successfully analyzed real samples of bees, larvae and bottom hive debris. Due to the high reliability and relatively simple detection scheme, the digital ULISA can pave the way for a new generation of digital immunoassays with a strong potential for commercialization.
Keywords:
single-molecule detection; photon-upconversion nanoparticle; bioconjugation
Available at various institutes of the ASCR
Photon-upconversion nanoparticles for single-molecule immunosensing of cancer biomarkers and bacteria
The recent progress in the field of immunoassays has been driven by introduction of various kinds of nanomaterials. In particular, photon-upconversion nanoparticles (UCNPs) proved to be excellent ...
Fast and highly sensitive laser scanner for recording photon-upconversion luminiscence from planar surfaces
Hlaváček, Antonín; Křivánková, Jana; Foret, František
2020 - English
Photon-upconversion nanoparticles (UCNPs) are lanthanide-doped nanocrystals that can be excited by nearinfrared light and emit photon-upconversion luminescence of shorter wavelengths. Advantages of UCNPs include near-infrared excitation, multiple and narrow emission bands, negligible autofluorescence and high stability, which make UCNPs ideal luminescence label for use in biological and chemical assays. These assays - e.g. upconversion-linked immunosorbent assay, western blot, lateral flow assay, gel electrophoresis, thin layer chromatography - commonly require the scanning of a planar surface with a high spatial resolution and an excellent sensitivity. The availability of commercial equipment is recently limited because of the novelty of the photon-upconversion phenomenon. Therefore, we report on the construction of photon-upconversion laser scanner. The scanner consists of a laser scanning head, which is attached to a xy-moving stage. The scanning head itself is constructed as an epiluminescence detector with excitation wavelength of 976 nm. A CCD array spectroscope is connected to the laser head and serves as a sensitive detector of photon-upconversion luminescence. The scanner possesses a spatial resolution of 200 μm, the scanning rate is up to 57 points per second and the sensitivity reaches down to single photon-upconversion nanoparticle.
Keywords:
photon-upconversion; nanoparticle; laser scanner; hyperspectral imaging
Available in digital repository of the ASCR
Fast and highly sensitive laser scanner for recording photon-upconversion luminiscence from planar surfaces
Photon-upconversion nanoparticles (UCNPs) are lanthanide-doped nanocrystals that can be excited by nearinfrared light and emit photon-upconversion luminescence of shorter wavelengths. Advantages of ...
Photon-upconversion scanner for multiplexed imaging
Hlaváček, Antonín; Křivánková, Jana; Foret, František
2020 - English
In analytical chemistry, multiplexed assays facilitate parallel detection of analytes. The multiplexing reduces the consumption of sample, reagents, and other resources and can be applied for medical diagnostics, the studies of biomolecule interactions, single-cell and singlemolecule assays, environmental monitoring, pathogen detection, multiparameter chemical and biological assays and screening of chemical libraries.
Keywords:
photon-upconversion scanner; photon-upconversion nanoparticles; droplet microfluidics
Fulltext is available at external website.
Photon-upconversion scanner for multiplexed imaging
In analytical chemistry, multiplexed assays facilitate parallel detection of analytes. The multiplexing reduces the consumption of sample, reagents, and other resources and can be applied for medical ...
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