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Filter switching device for dual-wavelength videoimaging
Authors:A N Chanturiya
Institution:(1) Laboratory of Theoretical and Physical Biology, National Institute of Child Health, National Institutes of Health, 20892 Bethesda, Maryland;(2) A. V. Palladin Institute of Biochemistry, 252030 Kiev, Ukraine;(3) LTPB/NICHD-NIH, Bd. 10, Rm. 10D04, 10 Center Drive, 20892 Bethesda, MD
Abstract:An inexpensive, dual-wavelength, videoimaging system that can be used for parallel observation of two fluorescent dyes is described. All four filters, two for excitation and two for emission, are placed on the same oscillating holder. Filters are coupled with a single dichroic mirror having two spectral windows. A coil driven by an electronic circuit connected to photosensors, which determine the position of the holder, moves the magnet that shifts the position of the filters. Since the filter holder is placed between two springs, it oscillates with the frequency of mechanical resonance. As a result the filter switching did not require much power and did not produce significant vibrations of the base. Switching frequencies up to 4.5 s–1 were reached with the first experimental device. System performance was tested using phospholipid vesicles loaded with water-soluble and membrane dyes. It has been demonstrated that the device can be used successfully in experiments on membrane fusion with rhodamine- and calcein-labeled liposomes.
Keywords:Fluorescence  microscopy  imaging  membrane fusion
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