Abstract 2094: Pathology beyond 2D: Imaging and analysis of cells and the tumor microenvironment in three dimensions

• Published in: Cancer research (Chicago, Ill.) Ill.), 2018-07, Vol.78 (13_Supplement), p.2094-2094
• Main Authors: Mansfield, James R., Wainright, James, Bagnall, Christopher, Wilde, Geraint, Price, Meredith, Willcox, Benjamin E., Browne, Mark
• Format: Article
• Language: English
• ISSN: 0008-5472; 1538-7445
• DOI: 10.1158/1538-7445.AM2018-2094

The success of cancer immunotherapy treatments and the development of immuno-oncology into the “fifth pillar” of cancer treatment has fueled a rapid growth in interest into methodologies that investigate immune cells in the context of the tumor microenvironment. However, nearly all technologies that investigate the immune contexture of tumors either rely on homogenization or disaggregation of the sample and the concomitant loss of spatial relationships, or rely on a thin section (5 um or less), two-dimensional sample of a biopsy, which is a poor representation of an inherently three-dimensional structure. An image of a thin section will tend to over-estimate the numbers of proximal cells and underestimate the numbers of more distant cells due to sampling error. To address this, we have implemented an imaging system capable of 3D confocal and super-resolution imaging, tiling of large sample areas and imaging samples over time along with software for the visualization and analysis of the datasets. We present here a series of examples showing the utility of 3D fluorescence imaging of labeled cells in thick (ca. 50 micron) FFPE tissue sections. By using a multimarker immunostaining scheme, this enables the phenotyping of cells within the tumor microenvironment and the calculation of distance metrics between cells in three dimensions. This methodology is an inherently better means of assessing cellular distributions in the tumor microenvironment and can give a much better representation of the complicated 3D structure of a tumor than imaging of thin sections. Citation Format: James R. Mansfield, James Wainright, Christopher Bagnall, Geraint Wilde, Meredith Price, Benjamin E. Willcox, Mark Browne. Pathology beyond 2D: Imaging and analysis of cells and the tumor microenvironment in three dimensions [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2094.