Spectral Imaging

We routinely use spectral imaging in our experiments. In this technique the emission spectrum is recorded for each an every pixel in the image. Applications include: verification of the labeling of specimens, for instance expression levels of fluorescent proteins, characterization of luminescent nano-particles and FRET. High sensitivity is achieved by using a prism for the dispersion in combination with a state-of-the-art back illuminated 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. The spectrograph is fiber-coupled to a commercial confocal laser scanning microscope (Nikon PCM2000).

Frederix, P., M.A.H. Asselbergs, W. Van Sark, D.J. Van den Heuvel, W. Hamelink, E.L. De Beer, and H.C. Gerritsen. 2001. High sensitivity spectrograph for use in fluorescence microscopy. Appl. Spectrosc. 55(8):1005-1012