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6th ESACP Congress, Heidelberg, April 7-11, 1999 |
A100
In modern pathology, assessment of diagnostic and prognostic molecular
markers has become increasingly important. It is conceivable that multi-
parameter diagnostic tests for phenotypic and genotypic markers will be
developed in the next few years. Assessment of morphological features remains
the golden standard in pathological diagnosis and, therefore, it is crucial
that molecular screening is implemented in a routine diagnostic environment,
using conventional bright-field microscopy. Multi-color bright- field
microscopy using enzyme precipitates has become feasible for
immunocytochemistry (IC) and in situ hybridization (ISH) techniques over the
past few years. Conventional bright-field microscopy optics have allowed up
to three targets to be detected simultaneously by the unaided eye. To apply
these methods reliably to pathological practice, unambiguous classification
of multi-color detection signals is required. Therefore, digital image
capture and color recognition computer programs has been introduced. Very
recently, Fourier-transform spectral imaging has been applied successfully to
fluorescence microscopy allowing unambiguous classification of 24-color
fluorescence ISH to human chromosomes (spectral karyotyping). In parallel to
spectral imaging of multi-color fluorescence, we explore the potential of
spectral analysis of multi- color bright-field microscopic specimens
(spectral pathology). Our data show unequivocal classification of a triple-
color ISH for chromosome centromeres in tumor cells, and accurate color
segmentation of immunostained "agarcyto" cell block specimens. Morphological
stains can be used simultaneously and are identified by color spectra. From
these promising data we conclude that spectral imaging has great potential
for qualitative and quantitative multi-parameter pathological diagnosis.
SPECTRAL PATHOLOGY: MULTICOLOR SPECTRAL IMAGING OF CHROMOGENETIC DYES IN
PATHOLOGICAL SPECIMENS
Macville M 1, Speel EJ 2, Hopman A 3, Van der Laak J 1,
Hanselaar A 1, McNamara G 4, Soenksen D 4, Ried Th 5
1 Pathology, University of Nijmegen, Netherlands, 2 Pathology, University
of Zuerich, Switzerland, 3 Cell Biology & Genetics, University of Maastricht,
Netherlands, 4 Applied Spectral Imaging, Inc., Carlsbad, CA, USA, 5 Division
of Clinical Sciences, NCI, NIH, Bethesda, MD, USA.