TY - JOUR
T1 - Spectral imaging of multi-color chromogenic dyes in pathological specimens
AU - Macville, Merryn V.E.
AU - Van Der Laak, Jeroen A.W.M.
AU - Speel, Ernst J.M.
AU - Katzir, Nir
AU - Garini, Yuval
AU - Soenksen, Dirk
AU - McNamara, George
AU - De Wilde, Peter C.M.
AU - Hanselaar, Antonius G.J.M.
AU - Hopman, Anton H.N.
AU - Ried, Thomas
PY - 2001
Y1 - 2001
N2 - We have investigated the use of spectral imaging for multicolor analysis of permanent cytochemical dyes and enzyme precipitates on cytopathological specimens. Spectral imaging is based on Fourier-transform spectroscopy and digital imaging. A pixel-by-pixel spectrum-based color classification is presented of single-, double-, and triple-color in situ hybridization for centromeric probes in T24 bladder cancer cells, and immunocytochemical staining of nuclear antigens Ki-67 and TP53 in paraffin-embedded cervical brush material (AgarCyto). The results demonstrate that spectral imaging unambiguously identifies three chromogenic dyes in a single bright-field microscopic specimen. Serial microscopic fields from the same specimen can be analyzed using a spectral reference library. We conclude that spectral imaging of multi-color chromogenic dyes is a reliable and robust method for pixel color recognition and classification. Our data further indicate that the use of spectral imaging (a) may increase the number of parameters studied simultaneously in pathological diagnosis, (b) may provide quantitative data (such as positive labeling indices) more accurately, and (c) may solve segmentation problems currently faced in automated screening of cell- and tissue specimens.
AB - We have investigated the use of spectral imaging for multicolor analysis of permanent cytochemical dyes and enzyme precipitates on cytopathological specimens. Spectral imaging is based on Fourier-transform spectroscopy and digital imaging. A pixel-by-pixel spectrum-based color classification is presented of single-, double-, and triple-color in situ hybridization for centromeric probes in T24 bladder cancer cells, and immunocytochemical staining of nuclear antigens Ki-67 and TP53 in paraffin-embedded cervical brush material (AgarCyto). The results demonstrate that spectral imaging unambiguously identifies three chromogenic dyes in a single bright-field microscopic specimen. Serial microscopic fields from the same specimen can be analyzed using a spectral reference library. We conclude that spectral imaging of multi-color chromogenic dyes is a reliable and robust method for pixel color recognition and classification. Our data further indicate that the use of spectral imaging (a) may increase the number of parameters studied simultaneously in pathological diagnosis, (b) may provide quantitative data (such as positive labeling indices) more accurately, and (c) may solve segmentation problems currently faced in automated screening of cell- and tissue specimens.
KW - Absorption
KW - Bright-field microscopy
KW - Fourier-transform spectroscopy
KW - In situ hybridization, immunocytochemistry
KW - Multicolor
KW - Optical density
KW - Pathology
KW - Spectral imaging
UR - http://www.scopus.com/inward/record.url?scp=0034919788&partnerID=8YFLogxK
U2 - 10.1155/2001/740909
DO - 10.1155/2001/740909
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C2 - 11455032
AN - SCOPUS:0034919788
SN - 0921-8912
VL - 22
SP - 133
EP - 142
JO - Analytical Cellular Pathology
JF - Analytical Cellular Pathology
IS - 3
ER -
