Flow cytometry and other single cell technologies 

Extensive experience with flow cytometry analysis and sorting of human and mouse lung, skin, hair follicle and thymus. I learned FACS during my PhD in the US, at University of Pittsburgh and at Duke University.  Flow cytometry was the entry point into the single cell world. Since then, I implemented single cell technologies both at EPFL (a Biomark, Fluidigm instrument) as well as in Giessen. In Giessen, we purchased several single cell instruments, we trained users and provided opportunities for continual training in the form of workshops, and single cell clubs. 

Human alveolar epithelial type II cells in healthy and IPF lung

What is an alveolar epithelial cells? Are there subtypes of AEC2s in the human lung? How are they replenished? And how does everything change in IPF. I started with flow cytometry characterization, based on known markers. Next I performed microarray transcriptomic analysis on sorted human AEC2s from normal and IPF lungs. I found new markers and identified new processes that improved our understanding of the human lung.

Precision cut lung slices

PCLS represent very thin slices (200-500 microns) of lung tissue that can be maintained in culture. They offer the advantage of having a quasi normal cellular environment that can be manipulated for the purpose of pharmaceutical drug discovery and development. This assay is very instrumental also in studying cell behavior during injury and repair, in healthy and diseased tissues. To confer the solid consistency necessary for sectioning, the lung has to be filled with low melt agarose, cooled on ice, and then sectioned. While not at all difficult to perform, organ procurement is a major concern. The technique needs to be adjusted for different diseases, according to the changes in tissue structure. Other challenges that we had to overcome were to establish culturing conditions and understand the cellular dynamics during the time of culture. This work led to several scientific collaborations and a patent.

Roxana Wasnick MD PhD 

Medical Scientist

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