Abstract

Imaging in 3 Dimensions; Paradigms and Pitfalls

Steven Titus, Staff Scientist, NIH - NCATS

The recent trend towards capturing and analyzing samples in three dimensions has increased as cell and disease models become more “physiologically relevant”. Many parameters need to be taken into consideration when initiating a 3D High Content Imaging and Analysis campaign. First, there are many samples such as single and multi cell type tumor spheroids, organoids, and organisms such as nematodes, fruit flies, and zebrafish that can be interrogated to explore different aspects of a disease or metabolic state. Second, many current and emerging imaging technologies such as widefield, confocal, deconvolution, and light sheet microscopy have evolved to acquire samples in 3D in an automated fashion. Third, there are many new plates such as micropatterned, hanging drop, and round bottom used to promote the growth of 3D cell models. And finally, practical considerations such as the loss of excitation and emission light in thick samples, celluolar penetration of UV to near IR imaging probes, and sample resolution also need to be taken into consideration before and during said campaigns. During this presentation, I will highlight the current and emerging trends in hardware; in terms of both microscopy and plate/sample types , imaging probes available, analysis and visualization software, and finish with some of the common pitfalls observed.