High sensitivity spectrograph for use in fluorescence microscopy
| Autor: | Frederix, P. L. T. M., Asselbergs, M. A. H., van Sark, W. G. J. H. M., van den Heuvel, D. J., Hamelink, W., de Beer, E. L., Gerritsen, H. C., Dep Natuurkunde, Section Energy and Resources, Sub Molecular Biophysics |
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| Přispěvatelé: | Dep Natuurkunde, Section Energy and Resources, Sub Molecular Biophysics |
| Jazyk: | angličtina |
| Rok vydání: | 2001 |
| Předmět: |
0301 basic medicine
medicine.medical_specialty Time resolved spectroscopy 03 medical and health sciences Optics Taverne Fluorescence microscope medicine Emission spectrum Spectral resolution Instrumentation Spectrograph Spectroscopy Fluorescence microscopy 030102 biochemistry & molecular biology business.industry Chemistry Quantum dot Spectral imaging Dwell time 030104 developmental biology Prism Time-resolved spectroscopy business IVMA |
| Zdroj: | Applied Spectroscopy, 55(8), 1005. Society for Applied Spectroscopy |
| ISSN: | 0003-7028 |
| DOI: | 10.1366/0003702011953153 |
| Popis: | In this paper a versatile, high sensitivity spectrograph is presented for use in fluorescence microscopy. The high sensitivity is achieved by using a prism for the dispersion in combination with a state-of-the-art back illuminated charge-coupled device (CCD) camera. The spectrograph, including the CCD camera, has a detection efficiency of 0.77 ± 0.05 at 633 nm. Full emission spectra with a 1–5 nm spectral resolution can be recorded at a maximum rate of 800 spectra per second. Two applications are shown, in which the spectrograph is fiber-coupled to a commercial confocal laser scanning microscope. In the first example, Förster resonance energy transfer imaging experiments were carried out on double-labeled actin filaments in the in vitro motility assay. A 160 × 160 point image was recorded in 1.5 min at 3 ms dwell time per image point. In the second application, a time-resolved study of single quantum dots is presented at 5.2 ms time resolution. |
| Databáze: | OpenAIRE |
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