S. cerevisiae is an excellent reference organism for biomedical research in investigating fundamental cellular processes. In spite of their small size, the cellular morphology and physiology of yeast cells can be successfully studied thanks to microscopy techniques such as indirect Immunofluorescence and fluorescent live cell imaging.
Indirect immunofluorescent protocols for specific and homogenous staining are typically easily applied, but at the cost of nonviable cells and time-consuming preparation of slides (procedure prone to problems with fixation and permeability). On the other hand, setting up live-cell, time-lapse microscopy requires expensive equipment and labeling with organelle-specific fluorescent dyes expression can be critically dependent on the yeast growth phase.
Nanolive’s technology wants to bypass these limits, allowing for a 3D live-cell imaging without any need for chemical markers or genetic manipulation. Based only on their specific Refractive Index (RI), all of the yeast’s different components can be distinguished and colored thanks to a process called digital staining. This allows the 3D Cell Explorer to visualize complex biological processes over time and in three dimensions, thus giving unprecedented insights into the dynamic cellular processes and physiology.
In this blog post we show how the 3D Cell Explorer is particular suited for:
- Real-time acquisitions of the budding phenomena (3 hours time-lapse, highest acquisition rate: 3D reconstruction in less than 2 seconds)
- Studying differences in bud and mother yeast (different composition, volume, RI, vacuole dimension etc.)
- Direct measures of cytoplasm/vacuole volume during the growing/starvation/aging phenomena
- Better morphological view of the yeast (localization of yeast and buds in three dimensions)
For the sample granting and the kind permission to reproduce the images on this blog post we thank Prof. Dr. Karsten Weis, Head of Institute of Biochemistry, ETH Zurich.