Lectures: 1.-5. Optical methods Features of electromagnetic radiation, interaction of radiation with matter. Radiation absorption and its principles and rules. Basic terminology and principles of absorption molecular spectrophotometry. Transmittance, absorbance, Lambert - Beer law, validity and verification of Lambert - Beer law. Absorption coefficient, absorption spectrum. Method of calibration curve, standard solutions, construction of calibration curve, calculation of calibration curve using linear regression. One standard method, calibration factor.Construction of machines for measuring of absorbance in the photometry and spectrophotometry. Sources of radiation energy, elements necessary for obtaining of monochromatic radiation. Absorption environment, detectors. Emission flame photometry, principle, flame photometry and its application in a clinical laboratory. Atomic absorption spectrophotometry. Turbidimetry. Nephelometry. Reflective photometry. Densitometry. Refractometry. Polarimetry. 6. Automatic analyzers and classification of them-spectrophotometric, electrophoretic, immunochemical. Laboratory information system. Equipment for the RIA. Luminometer. 7. Osmometry and oncometry.Osmotic pressure, oncotic pressure, osmolality, osmolality measurement and calculation. 8.-9. Electrochemical methods.Faraday's laws. Coulometric titration. Conductometry.Potentiometry. Polarographic methods. 10.Enzyme immunoanalysis. Quality control (external and internal) control in a laboratory. 11.-14. Chromatographic methods:Principle of chromatographic methods, classification of chromatography according to a physical principle, design, forces and effects applied in chromatography. Chromatography on paper. Thin layer chromatography. Gas chromatography. Liquid chromatography. Gel liquid chromatography. Ion-exchange liquid chromatography. Proper laboratory work.Internal and external quality control of laboratory work. 15. Electrophoretic methods.Principles of electrophoretic methods, isoelectric point, application in a clinical laboratory. Electrophoresis on cellulose acetate sheets, agar, agarose, polyacrylamide gels, capillary electrophoresis, application in a clinical laboratory.Isoelectric focusing, isotachophoresis, equipment, evaluation application in a clinical laboratory. Laboratory practicals: 1.-2. Demonstration of basic physical principles used in laboratory technique.Spectrophotometer-design (radiation sources, monochromators, interference filters, detectors). Flame-emission photometer - design, dosing of analytic sample.pH-met.Coulometer.Conductometer.Densitometer. 3.-15. Application of physical principles in laboratory technique. Spectrophotometer UV / VIS fluorimeter, nephelometers, densitometer, refractometer, reflective photometer, emission flame photometer, atomic absorption spectrophotometer. PH meter, polarograph, Coulometer, conductometer. Equipment for electrophoresis (agarose, polyacrylamide, isoelectric focusing).Equipment for thin-layer chromatography, liquid chromatography - HPLC, gas chromatograph, detectors. Osmometr.ISE (Na, K, Cl, Ca, Mg).Clarck´s oxygen electrode Severinghans´s pO2 electrode, enzyme electrodes. Centrifuges.Thermostats, water baths.
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A student is able to manipulate and use laboratory equipment. He/she acquires knowledge of calibration in accordance with proper laboratory work.Standard operating technical procedures (SOPT), instructions for operation and maintenance of equipment, operational diaries.Automatic analyzers (Abbott, Beckmann, Roche, Olympus).Communication with automated analytical systems, report on system status, error messages and reports. Check of an activity and setting of individual components of an analyzer (photometer check, ISE check, blank cell, setting pipettors.)Elements of daily, weekly and monthly maintenance of an analyzer.UV / VIS spectrophotometer, flame photometer emission and atomic absorption spectrophotometer. Osmometr.Analyzer of AB metabolism and blood gases (pH, pO2, pCO2) hemoximetr.Equipment for protein electrophoresis (manual and automatic version), densitometer.Liquid chromatograph, gas chromatograph.Flow cytometer.Coagulometers.Separators. Centrifuges.Thermostats, incubation bath.Equipment for microwave breakdown-mineralization of biological samples. |
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Dastych M. Instrumentální technika-obor zdravotní laborant. Masarykova univerzita 1. vydání 2007. ISBN 978-80-210-4226-1.
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Drechslerová, Taťána; Bečvářová, Ilona; Plačková, Tereza. Laboratorní technika. Vydání. 1. 2015. ISBN 978-80-261-0533-6.
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Hrazdira I., Mornstein V. Lékařská biofyzika a přístrojová technika, 1. vydání Brno: Neptun, 2001.
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Lewandrowski Kent. Clinical Chemistry, laboratory management-clinical correlations, Lippincott Williams &Wilkins, USA, 2002.
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Zima, T., Šebesta, I., Štern, P. Laboratorní technika I. a II. díl. Univerzita Karlova Praha, 1. lékařská fakulta 2005.
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