1) How did you compose this image?
The image of immortalized retinal pigment epithelial (RPE) cells migrating through a collagen matrix was prepared as a maximum intensity projection using Fiji (ImageJ). To more clearly reveal the structure of the vimentin (magenta) and microtubule (green) networks, the image was inverted.
2) How does this image reflect your scientific research?
We are interested in understanding how cells sense their environment and how this process influences cell architecture and signaling. Axially Swept Light-Sheet Microscopy (ASLM) allows us to simultaneously image sub-cellular and large-scale (~200 m) environmental features with isotropic high-resolution (~390 nm). As such, we can begin to address specific biological questions involving the multiple length scales present in this image. For example, what is the role of the vimentin network (a major marker of the epithelial to mesenchymal transition in cancer) in cell polarization and migration? What leads to the disassembly of the vimentin network during mitosis, and can this process be pharmacologically targeted? The cells featured in this image reveal many interesting phenotypes, each of which reflects scientific research opportunities moving forward.
3) Can you please provide a few real-world examples of your research?
Understanding physiological and pathophysiological processes in 3D microenvironments is fundamentally important in human development, health, and disease. In particular, it has major implications in cancer metastasis, where cells must shed cell-cell junctions, polarize, migrate away from the primary tumor, evade the immune system, intravasate into the bloodstream, extravasate, colonize a secondary tissue, and proliferate. By studying aspects of these processes with molecular precision in reconstituted tissue-like environments, we hope to make a lasting impact that directly leads to improved patient outcomes.
4) How does your research apply to those who are not working in your specific field?
ASLM is an enabling imaging technology that may be applied to diverse scientific disciplines. Most immediately, we see significant opportunities in cellular and developmental biology, where ASLM allows researchers to visualize subcellular structures in large, mechanically unperturbed 3D samples without spatial bias. This capability is invaluable because of the effects that the extracellular microenvironment plays on cell architecture, cell signaling, and cell fate. Alternatively, ASLM could be used by neuroscientists measuring the ‘connectome,’ where near-diffraction-limited synaptic features must be resolved throughout large expanses of brain tissue.
5) Do you have a website where our readers can view your recent research?
– Kevin Dean, Erik Welf, Gaudenz Danuser, and Reto Fiolka