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Improved high-content screening of yeast mutant collections
We have improved our screening approach for imaging fluorescently labeled yeast strain collections (1,2,3):
* Imaging of yeast cells immobilized as a monolayer in 386well microtiter plates.
* High-resolution, true 3D multi-channel imaging.
* Reliable auto-focus system.
* Imaging both of living and fixed cells.
* Acquisition of a large number of cells at once.
* ~90% reliability
* Automated image processing (HCS Profiler, CellSerpent or other software).
* Data storage using YPL+.
The screening approach (Wolinski et al. in prep.) is compatible with SGA technology and our new CARS (coherent anti-Stokes Raman Scattering) system..
If you are interested in collaborations please contact:
1. Wolinski H, Petrovic U, Mattiazzi M, Petschnigg J, Heise B, Natter K & Kohlwein SD. 2009. Imaging-based live cell yeast screen identifies novel factors involved in peroxisome assembly. J Prot Res 8(1):20-27.
2. Wolinski H, Natter K & Kohlwein SD. 2009. The fidgety yeast: focus on high-resolution live yeast cell microscopy. in Methods Mol Biol 548:75-99.
3. Wolinski Heimo, Kristian Bredies, Sepp D. Kohlwein 2011.Quantitative imaging of lipid metabolism in yeast: from 4D analysis to high content screens of mutant libraries. Methods in Cell Biology 108:345-65.
4. Tavassoli S, Chao JT, Loewen C. A high-throughput method to globally study the organelle morphology in S. cerevisiae.J Vis Exp. 2009 Mar 2;(25). pii: 1224. doi: 10.3791/1224.
5. A.H. Tong, M. Evangelista, A.B. Parsons, H. Xu, G.D. Bader, N. Page, M. Robinson, S.Raghibizadeh, C.W. Hogue, H. Bussey, B. Andrews, M. Tyers, C. Boone, Systematic genetic analysis with ordered arrays of yeast deletion mutants, Science 294 (2001) 2364-
6. Vizeacoumar FJ, van Dyk N, S Vizeacoumar F, Cheung V, Li J,Sydorskyy Y, Case N, Li Z, Datti A, Nislow C, Raught B, Zhang Z, Frey B, Bloom K, Boone C, Andrews BJ.Integrating high-throughput genetic interaction mapping and high-content screening to explore yeast spindle morphogenesis. Cell Biol. 2010 Jan 11;188(1):69-81.
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