TY - JOUR
T1 - High sensitivity spectrograph for use in fluorescence microscopy
AU - Frederix, P. L. T. M.
AU - Asselbergs, M. A. H.
AU - van Sark, W. G. J. H. M.
AU - van den Heuvel, D. J.
AU - Hamelink, W.
AU - de Beer, E. L.
AU - Gerritsen, H. C.
PY - 2001/8
Y1 - 2001/8
N2 - 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.
AB - 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.
KW - Fluorescence microscopy
KW - Spectral imaging
KW - Time resolved spectroscopy
KW - IVMA
KW - Quantum dot
U2 - 10.1366/0003702011953153
DO - 10.1366/0003702011953153
M3 - Article
SN - 0003-7028
VL - 55
SP - 1005
EP - 1012
JO - Applied Spectroscopy
JF - Applied Spectroscopy
IS - 8
ER -