Light sheet microscopy/SPIM is a promising technology for high temporal and spatial resolution, deep tissue and long-term live imaging. It becomes a well-established method to image scattering sample such as Zebrafish, Drosophila. Recently, we reported a major breakthrough in 3D MultiCellular Tumor Spheroids (MCTS) imaging based on the engineering of spheroids expressing a histone-H2B fluorescent nuclear reporter protein, and specifically designed sample holders to monitor live cell division dynamics in 3D large MCTS using the SPIM technology. Though SPIM imaging opens new perspectives in the understanding of biological processes, it still suffers from various specific defects. Light -scattering and -absorption in dense biological samples lead to the attenuation of the light sheet illumination, resulting in a progressive degradation in image quality and signal strength along the x- and z-axes. As samples are illuminated from the side using directed light, SPIM images are often impaired by stripes and shadows. Moreover, spatial variations in the specimen refractive index cause major changes to the light path, resulting in aberrant images. Aberration effects can also introduce geometric image distortions, degrade image resolution and reduce the signal-to-noise ratio. These effects are particularly strong when thick biological opaque specimens, such as large MCTS, are investigated. We will present an overview of our most recent technical advances and their applications in the imaging of tumor spheroids using light sheet microscopy.


Monday, April 22, 2013, 12:00. Seminar Room

Hosted by Prof. Pablo Loza-Álvarez