the metpiki67 of proliferation index estimation in samples ... · 3,3'-diaminobenzidine and...
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3 – Warsaw University of Technology, Warsaw, Poland 4 – Central Clinical Hospital of the Ministry of the Interior and Administration, Warsaw, Poland.
MATERIALS AND METHODS
RESULTS
DISCUSSION AND CONCLUSIONS
Anna Korzyńska1, Łukasz Roszkowiak1, Jakub Żak1, Dorota Pijanowska1, Krzysztof Siemion1,4,
Wojciech Kozłowski2, Renata Maryniak2, Tomasz Markiewicz2,3
BACKGROUND
Methods: The MetpiKi67 (METhod of Proliferation Index of Ki67) method quantifies proliferating
cells' index in samples from Diffuse Large B-cell Lymphoma (DLBCL) patients. This method
quantifies the percentage of immunopositive nuclei (proliferating cancer cells nuclei) to the whole
number of nuclei in selected region.
The proliferation index is the most important feature used in benign and malignant diagnosis. It quantifies the percentage of immunopositive nuclei (proliferating cancer cells nuclei) to the whole number of nuclei in selected region with Ki67 stains. The localization of the antigens are visualized with 3,3'-Diaminobenzidine and Haematoxylin (DAB&H). The automated quantification of this index is dependent of the architecture of tissue section and type of diseases what results in several such algorithms developed so far [1-4]. In the case of this study the method is adjusted to the samples from the patients diagnosed towards Diffuse Large B-cell Lymphoma (DLBCL), that is very aggressive and can cause high mortality. The samples are mostly biopsies of various lymph nodes, but also skin, tonsils, breast and so on. The lymph node architecture is not always present in samples. The B-cell nuclei are varying in the shape, size and its chromatin is arranged differently: sometimes is uniformly distributed, but mostly is distributed peripherally with dark brown spots on brighter brown background or opposite. The samples of lymph node and affected tissue in various magnification are presented below.
Proposed MetpiKi67 method performs well
according to visual inspection but it should be
evaluated in comparison to the gold standard.
We plan to compare the results with other
automatic methods [1-4] and with results of
human evaluation based on objects counting.
The higher the cancer grade the less homogeneous nuclei structures are observed. Their size is increasing and shape is varying. This essentially complicates the identification of individual portions of chromatin and combining them into one object.
Acknowledgements:
References:
This study was supported by the National Centre
for Research and Development, Poland
(grant PBS2/A9/21/2013).
Automated evaluation of the region
PI = 93% and
objects area ratio = 0.90
Pathologist evaluation of WSI = 90%
Automated evaluation of the region
PI = 80% and
objects area ratio = 0.73
Pathologist evaluation of WSI = 60%
Automated evaluation of the region
PI = 70% and
objects area ratio = 0.88
Pathologist evaluation of WSI = 90%
Automated evaluation of the region PI =
70% and
objects area ratio = 0.64
Pathologist evaluation of WSI = 90%
Automated evaluation of the region PI
= 89% and
objects area ratio = 0.95
Pathologist evaluation of WSI = 90%
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11 digital slides from Pathology Department of the
Military Institute of Medicine in the form of .mrxs files
(scanned by 3DHistech slide scanners) with diagnosis
and proliferation index estimated by experienced
pathologist was used to verify proposed method.
Skin of the left mental
region.
Lymph node of the right axillary
region.
The MetpiKi67 of Proliferation Index estimation in samples
from patients with Diffuse Large B-cell Lymphoma
1 – Nalecz Institute of Biocybernetics and Biomedical Engineering PAS, Warsaw, Poland. 2 – Military Institute of Medicine, Warsaw, Poland.
It is complicated to segment nuclei with peripheral chromatin than to select nuclei of cell not affected by cancer.