Digital Staining of Unstained Pathological Tissue Samples through Spectral Transmittance Classification

University of Pittsburgh, Pittsburgh, Pennsylvania, United States
Optical Review (Impact Factor: 0.66). 01/2005; 12(1):7-14. DOI: 10.1007/s10043-005-0007-0

ABSTRACT Histological structures of a pathological tissue sample convey information relevant to the diagnosis of the disease that might have afficted the person. To reveal the morphology of these structures clearly, pathological tissues are stained. In this paper, a digital staining methodology for pathological tissue samples is introduced. Digital staining implies the application of digital processing techniques to transform the image of an unstained sample to its stained image counterpart. In the method, the transmittance spectra of the unstained and Hematoxylin and Eosin (H&E) stained multispectral images (16 bands) of specific tissue components are utilized. Two experiments were conducted to probe the possibility of the digital staining framework: the linear mapping of spectral transmittances, and the classification of spectral transmittances in conjunction with the linear mapping of specific transmittance data sets. The method classified the four tissue components, e.g. nucleus, cytoplasm, red blood cells, and the white region (region devoid of tissue structures), while the misclassifications between components with spectral transmittances that are closely similar were not completely rectified. 2005 The Optical Society of Japan

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Available from: Masahiro Yamaguchi, Sep 27, 2015
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    • "Digital images allow the development of digital algorithms for tissue analysis,[4–7] hence are obvious candidates for computational analysis. The practical application of multispectral and hyper-spectral imaging to pathology has also attracted the attention of several researchers, particularly its usefulness in bringing out details that are otherwise inconspicuous with the conventional RGB color imaging.[8–10] "
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    ABSTRACT: The objective of this paper is to improve the visualization and detection of tissue folds, which are prominent among tissue slides, from the pre-scan image of a whole slide image by introducing a color enhancement method that enables the differentiation between fold and non-fold image pixels. The weighted difference between the color saturation and luminance of the image pixels is used as shifting factor to the original RGB color of the image. Application of the enhancement method to hematoxylin and eosin (H&E) stained images improves the visualization of tissue folds regardless of the colorimetric variations in the images. Detection of tissue folds after application of the enhancement also improves but the presence of nuclei, which are also stained dark like the folds, was found to sometimes affect the detection accuracy. The presence of tissue artifacts could affect the quality of whole slide images, especially that whole slide scanners select the focus points from the pre-scan image wherein the artifacts are indistinguishable from real tissue area. We have a presented in this paper an enhancement scheme that improves the visualization and detection of tissue folds from pre-scan images. Since the method works on the simulated pre-scan images its integration to the actual whole slide imaging process should also be possible.
    11/2010; 1(1):25. DOI:10.4103/2153-3539.73320
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    ABSTRACT: Conventional widefield light microscopy and confocal scanning microscopy have been indispensable for pathology and drug discovery research. Clinical specimens from diseased tissues are examined, new drug candidates are tested on drug targets, and the morphological and molecular biological changes of cells and tissues are observed. High throughput screening of drug candidates requires highly efficient screening instruments. A standard bio- medical slide is 1 by 3 inches (25.4 by 76.2 mm) in size. A typical tissue specimen is 10 mm in diameter. To form a high resolution image of the entire specimen, a conventional widefield light microscope must acquire a large number of small images of the specimen, and then tile them together, which is tedious, inefficient and error-prone. A patented new wide field-of-view confocal scanning laser imaging system has been developed for tissue imaging, which is capable of imaging an entire microscope slide without tiling. It is capable of operating in brightfield, reflection and epi-fluorescence imaging modes. Three (red, green and blue (RGB)) lasers are used to produce brightfield and reflection images, and to excite various fluorophores. This new confocal system makes examination of large biomedical specimens more efficient, and makes fluorescence examination of large specimens possible for the first time without tiling. Description of the new confocal technology and applications of the imaging system in pathology and drug discovery research, for example, imaging large tissue specimens, tissue microarrays, and zebrafish sections, are reported in this paper.
    Proceedings of SPIE - The International Society for Optical Engineering 11/2005; 6009. DOI:10.1117/12.630896 · 0.20 Impact Factor
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