A method and apparatus for an improved spectral imaging system is
provided. The system is capable of measuring the fluorescence,
luminescence, or absorption at selected locations on a sample plate. The
emissions detection subassembly can tune to any wavelength within a
continuum of wavelengths utilizing an interferometric spectral
discriminator. The interferometric spectral discriminator creates an
interferogram from which the wavelength spectra for each pixel of the
array can be calculated, typically using Fourier transform analysis. In
one aspect, the chromatic accuracy of the system is calibrated using a
calibration slit placed in the input aperture of the input relay lens but
outside of the sample image. The slit is illuminated using a source of
known wavelength. The fringe count versus the wavelength of the slit
illumination source is monitored and used to calibrate the spectral
discriminator. In another aspect, a transparent optic is included in the
interferometric spectral discriminator that can be inserted into the beam
path whenever a monochrome image of the sample is required. The optic
produces a large offset in the legs of the interferometer resulting in
the fringe density becoming too large to resolve by the individual pixels
of the detector array. In another aspect, the interferometric spectral
discriminator includes a polarizing beam splitter. The polarizing beam
splitter preferentially reflects one polarization while preferentially
transmitting a second polarization, thus achieving improved efficiency
while minimizing ghosting. In another aspect, a metaphase finder is used
to locate areas of interest. The sample plate containing the material of
interest is illuminated with light of a wavelength determined to
preferentially scatter from objects the size of the metaphase spreads.
The intensity of the scattered light versus the location on the sample
plate is monitored and used to locate the areas of interest. Preferably
the sample plate is also illuminated by light of a second wavelength
which is not preferentially scattered by the objects of interest, thus
representing the background scatter. By subtracting the background
scatter from the primary scattered light, improved object discrimination
is achieved.