Seminars at ASCB | EMBO 2018

December 9, 2018 – 12:00 PM - 12:30 PM | Theater 2, Learning Center

Celldiscoverer 7 and 3D-Cell Culture - High Resolution meets Screening

Dr. René Buschow
Max Planck Institute for Molecular Genetics, Berlin, Germany

The use of three-dimensional cell culture systems gain more and more attention in the field of developmental biology, personalized medicine and cancer research, as well as general cell biology. We developed an experimental pipeline that enables us to monitor and quantify the viabilities of 3D cell systems under live cell conditions.

Using a low magnification, i.e. large object fields, ultra-low attachment tissue cultures were imaged in 384-well plates in a fully automated mode. Downstream, the image data was transferred into an automated analysis and archiving pipeline (Columbus, Perkin Elmer).

Prerequisite for live cell imaging in 384-well-formats is a stable environmental control as well as a light efficient microscope to reduce exposure times, phototoxicity, and guarantees an adequate throughput. Further, our set up finally required a fluorescence 3D imaging routine for evaluation of fluorescence reporters under live cell conditions (e.g. marker, viability staining, or drug availability).

The hardware to encounter these experimental tasks is all combined within the novel microscopy platform Celldiscoverer 7 (by Zeiss). This fully automated research imaging system enables a broad range of magnifications (2.5x-100x), large working distances and full environmental control, making it the ideal instrument for live cell 3D-systems. Further, automated focus strategies and outstanding Autocorr objectives ensure optimal imaging and constant photon yields for a reliable quantification of fluorescence signals in combination with modern 3D-deconvolution algorithms.

We use ZEN 2.5 as imaging and deconvolution software, for initial processing steps and established a data transfer pipeline. The final image analysis and archiving is performed using PerkinElmers Columbus platform.

The combination of the superior optics and imaging flexibility of the Celldiscoverer 7 plus the analysis platform Columbus (Perkin Elmer) enables the hybrid between modern High Content Microscopy with the flexibility required in academic research.