
Proceedings Paper
Quantification of relative chromatin content in flow cytometry standards using 3D OPTM imaging techniqueFormat | Member Price | Non-Member Price |
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Paper Abstract
A potential biomarker for early diagnosis of cancer is assessment of high nuclear DNA content. Conventional
hematoxylin staining is neither stoichiometric nor reproducible. Although feulgen stain is stoichiometric, it is time
consuming and destroys nuclear morphology. We used acidic thionin stain, which can be stoichiometric and also
preserve the nuclear morphology used in conventional cytology. Fifty chicken erythrocyte nuclei singlets (CENs), diploid trout erythrocyte nuclei (TENs) and Triploid TENs were stained for 15 and 30 minutes each. After imaging with optical projection tomography microscope (OPTM), 3D reconstructions of the nuclei were processed to calculate
chromatin content. The mean of ratios of individual observations was compared with standard ratios of DNA indices
of the flow cytometry standards. Mean error, standard deviation and 97% confidence interval (CI) was computed for
the ratios of these standards. At 15 and 30 minutes, the ratio of Triploid TEN to TEN was 1.72 and 1.76, TEN to CEN
was 1.27 and 2.01 and Triploid TEN to CEN was 2.11 and 3.39 respectively. Estimates of DNA indices for all 3 types
of nuclei had less mean error at 30 minutes of staining; Triploid TEN to TEN 0.349±0.04, TEN to CEN 0.36±0.04 and Triploid TEN to CEN 0.64 ± 0.07. In conclusion, imaging of cells with thionin staining at 30 minutes and 3D reconstruction provides quantitative assessment of cell chromatin content. The addition of this quantitative feature of aneuploidy is expected to add greater accuracy to a classifier for early diagnosis of cancer based on 3D cytological imaging.
Paper Details
Date Published: 29 March 2013
PDF: 11 pages
Proc. SPIE 8676, Medical Imaging 2013: Digital Pathology, 86760T (29 March 2013); doi: 10.1117/12.2007281
Published in SPIE Proceedings Vol. 8676:
Medical Imaging 2013: Digital Pathology
Metin N. Gurcan; Anant Madabhushi, Editor(s)
PDF: 11 pages
Proc. SPIE 8676, Medical Imaging 2013: Digital Pathology, 86760T (29 March 2013); doi: 10.1117/12.2007281
Show Author Affiliations
Nitin Agarwal, Univ. of Washington (United States)
Alberto M. Biancardi, Cornell Univ. (United States)
Florence W. Patten, VisionGate, Inc. (United States)
Alberto M. Biancardi, Cornell Univ. (United States)
Florence W. Patten, VisionGate, Inc. (United States)
Published in SPIE Proceedings Vol. 8676:
Medical Imaging 2013: Digital Pathology
Metin N. Gurcan; Anant Madabhushi, Editor(s)
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