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44 Bioanalytics 5.3

44 Bioanalytics 5.3 Bioanalytics Introduction The accurate analysis of chemical compounds, biological macromolecules as well as cells and microorganisms is currently an important problem being investigated in a variety of scientific, industrial and medical fields. Microscopic and spectroscopic procedures are the technologies commonly used in these investigations. Favorite approaches, especially with a number of biological specimens, are high-performance liquid chromatography (HPLC) with UV or fluorescence detectors and Raman spectroscopy. Berlin and Brandenburg have developed an industrial or industry-oriented infrastructure that makes available technologies for the analysis of biological compounds as well as other devices. Problems such as diagnosing skin cancer demonstrate how biological diagnostics can bring about advances in clinical diagnostics. Fluorescence and Raman Spectroscopy Mobile analysis technology for fluorescence and Raman spectroscopy is being developed at the Fraunhofer Institute for Reliability and Microintegration (Fraunhofer IZM). By virtue of the variety of radiation wavelengths it employs, the RF-KombiSCAN not only can detect the quantity of different substances, but their composition as well. This innovative, portable, hand-held optical meter simplifies and expedites the process of making measurements through the integration of fluorescence and Raman spectroscopy. The mobile scanner can be used as a laboratory research device as well as in industrial applications. The technical innovation and mobility of the RF-KombiSCAN is currently unsurpassed and could be applied to other areas as well, such as food monitoring, medical sciences, police investigations, border protection measures, forensics as well as in inorganic material analysis. The Ferdinand-Braun-Institut, Leibniz-Institut für Höchstfrequenztechnik (FBH) develops laser emitters for Ra- man spectroscopy. This includes diode lasers that emit light with two wavelengths at a fixed distance of about 1 nm from a single chip, which makes them ideal for “shiftedexcitation Raman difference spectroscopy” (SERDS). SERDS makes it possible to clearly separate Raman signals from interfering background fluorescence or ambient light. Use of micro-optics means the laser can be kept compact, which is essential for mobile use. FBH uses this technology in order, among other things, to provide beam sources in the yellow wavelength range. RF-KombiSCAN as laboratory device © Fraunhofer IZM Miniaturized two wavelengths diode laser module for Raman spectroscopy © FBH/Schurian

Bioanalytics 45 Imaging Raman Spectroscopy Imaging Raman spectrometers are multichannel (“multiplex”) devices that are not yet available as industrial products. Due to the extremely high demands placed on modern large telescopes, imaging multiplex spectrographs have existed in astrophysics for about 25 years. The goal of the RIA project being carried out jointly by the Laserund Medizin-Technologie GmbH, Berlin (LMTB), the Leibniz Institute for Astrophysics Potsdam (AIP) and the companies Berliner Glas and eagleyard photonics is to develop such a multichannel Raman spectroscopy system. The following areas were addressed as pilot applications for this project: • Analysis in microtiterplates: endpoint and kinetics in microtiter • Process analytics/recycling: identification of substances and their transformation during processing. • Security: identifying substances and liquids at access control, e.g. at airports. | | Fluorescence In Vivo Imaging The principle of fluorescence in vivo imaging is based on the properties of fluorochromes, which respond to an external light source by emitting light of a different wavelength. greateyes supplies a camera capable of detecting in the near-infrared range that is able capture the emissions. Fluorescence in vivo imaging can be applied, for example, to detect cancer cells in lymph nodes. In the example above, a fluorescence labelled dye was applied intravenously into rats, which then concentrated in lymph node tissues. The detection of the weak fluorescence, which permeates the tissue, requires a highly sensitive camera together with a special filter. Time-resolved Optical Microscopy Studies in bioenergetics in the Department of Chemistry at TU Berlin employ microscopy and spectroscopy and electrophysiological methodologies, combining them with optical methodologies in examining living cells. Wide-field fluorescence microscopy with high spacial- and time-resolution single photon detectors for multichannel FLIM measurements provide for spatial resolution microscopy of dynamic processes and simultaneous fluorescence correlation spectroscopy in each pixel with 100 ps resolution along with a measurement period of 10 microseconds. These technologies go beyond previous limits in terms of precision, parallelization and speed and have a particularly high potential for applications in industrial projects. This applies in particular to applications in the field of pharmaceutical active ingredient research and cell-based diagnostics. This makes it possible, for example, to detect with unprecedented precision the fluorescence lifetime distribution of photosynthetically active proteins in unicellular organisms in order to investigate important regulatory processes in photosynthesis. Biological or chemical reference structures will help in determining to what extent the technology can overcome the diffraction limit of spatial resolution in optical microscopy procedures. Overlay picture of a light scatter image (left) and fluorescence detection (right) using the camera GE 1024 1024 DD NIR © greateyes GmbH

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