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Quantitative Measurement of Apoptotic and Mitotic Nuclear Events Using Automated Classification of Bright Field and Nuclear Fluorescent Imagery with the ImageStream MultiSpecral Imaging Flow Cytometer
Author(s) -
Morrissey Philip,
Henery Shan,
George Thaddeus,
Venkatachalam Vidya,
Wardhani Aster,
Ortyn William,
Basiji David
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.1238.16
Subject(s) - mitosis , mitotic index , multispectral image , artificial intelligence , biology , computer science , microbiology and biotechnology
Both apoptotic and mitotic cells exhibit nuclear condensation which can be measured in a quantitative manner using a simple DNA dye and the ImageStream multispectral imaging flow cytometry. To distinguish apoptotic from mitotic cells additional morphological measurements such as textured brightfield imagery and increased texture and intensity of dark field scatter imagery may also be required. Distinguishing the sometimes minimal difference in level of nuclear condensation between apoptotic and mitotic cells in an automated manner can be problematic when applying only individual intensity and texture image analysis parameters. Here we present an automatic classifier which generates complex weighted combinations of intensity and texture features applied to multiple images of each cell to optimize distinction and quantitation of apoptotic and mitotic events in the same sample. Imagery from Ramos, Jurkat, and Colo205 cell lines were acquired on the ImageStream multispectral imaging flow cytometer after incubation with mechanistically distinct apoptosis inducing drugs (Camptothecin and Staurosporine) and staining with simple nuclear dyes. The accuracy of an automatic classifier which can measure the apoptotic index and mitotic index of samples based on brightfield, nuclear, and darkfield scatter imagery without the need for additional fluorescent labels using the IDEAS analytical software package is demonstrated.