Paul Vigneau Jr
Research Mentor: Marisa Aikins
Mentor Department: Internal Medicine, Medicine
Author(s): Paul Vigneau, Andrew Yu, Tejas Thiyagarajan, Sydney Malawer, Madeline Bough, Marisa Aikins, Sofia Merajver
Session: Session 6 (3:00 PM – 3:50 PM)
Presentation Type: Poster 99
Abstract
The goal of this research is to develop a methodology that can be used to image large and dense ex vivo tumor organoids (ALK+ NSCLC organoids harvested from mice as proof of concept). To accomplish this a light sheet microscopy methodology has been employed to spatially visualize immune cells within the tumor organoid. Currently, the major obstacles to obtaining a clear image are autofluorescence, fluorescence preservation, and issues with the interaction between the DAPI and green nuclear stains (used to label spheroid and immune cells, respectively). Methods used to address these obstacles include the testing of different fluorescent stains such as a long-term cytoplasmic, optimizing staining protocol via incubation times and stain diffusion rate, and adjusting the clearing method and subsequent light sheet laser power and acquisition order accordingly. It is expected that by optimizing the type of stain used to label the immune cells it will be possible to differentiate between immune cells and nonimmune cells using this process. If true, the resolution of cells will be clear, and the methodology will be able to reliably determine spatial localization and frequency of immune cells within the organoids through a downstream computational analysis pipeline.
