Abstract
Next-generation 3D high-content imaging and analysis enables the quantification and display of tissue heterogeneity regarding subcellular properties, and hence will improve the diagnostic characterization of biopsied tissues. ( Upper left) Needle-biopsy section of human prostate adenocarcinoma, immuno-labeled for global DNA methylation (5methylcytosine, false-colored green) and counterstained for global nuclear DNA with 4',6-diamidino-2-phenylindole (DAPI, false-colored blue) and imaged with a 20X NA=0.7 lens.
Highlights
3D reconstruction of a stack of confocal images taken from the boxed region
processed using maximum intensity projection combined with shadow projection
of cells according to the levels of global nuclear DNA methylation using dedicated software
Summary
Next-generation 3D high-content imaging and analysis enables the quantification and display of tissue heterogeneity regarding subcellular properties, and will improve the diagnostic characterization of biopsied tissues. (Upper left) Needle-biopsy section of human prostate adenocarcinoma, immuno-labeled for global DNA methylation (5methylcytosine, false-colored green) and counterstained for global nuclear DNA with 4',6-diamidino-2-phenylindole (DAPI, false-colored blue) and imaged with a 20X NA=0.7 lens. (Upper right) 3D reconstruction of a stack of confocal images taken from the boxed region (left sub-image with a 63X NA=1.3 lens, and processed using maximum intensity projection combined with shadow projection (CytoFx software, llucida LLC). (Lower left) Classification (phenotyping) of cells according to the levels of global nuclear DNA methylation using dedicated software; high (cyan) and mid levels (red) found predominantly in the epithelial-ductal cells, and low levels (blue) in stromal cells. (Lower right) 3D visualization of the classified heterogeneous cell population. Translational Cytomics Group, Cedars-Sinai Medical Center, Los Angeles, CA, USA *Corresponding author: Jian Tajbakhsh, Translational Cytomics Group, Cedars-Sinai Medical Center, Los Angeles, CA, USA, Tel: +1 310-423-3277; E-mail: tajbakhshj@cshs.org
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