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Equipment
Yeast facility
A dedicated room for using yeast as a model system has been setup, comprising all the standard equipment for handling yeast cultures. This includes several dry incubators, three shaking dry incubators, two dissection microscopes for tetrad analysis, a PSM for media plates preparation, and a clinical centrifuge. This core facility is open to all yeast users of the Institute.
Microscopy
  • Spinning disk confocal microscope
  • Epifluorescence microscope
  • Routine
    microscope
  • Image
    analysis
For live-cell imaging, we are equipped with a complete state-of-the-art spinning disk confocal system integrating the new wide-field CSU-W1 head from Yokogowa, which provides us with better confocality and takes advantage of the wide field-of-view of the new sCMOS cameras. It is mounted on an inverted microscope operating with a hardware autofocus system and a piezoelectric stage for fast and accurate Z movements. In addition, the motorized bypass of the disk allows for epifluorescence imaging using a LED light source. This microscope is dedicated to live imaging of fission yeast cells grown in microfluidic devices.
    Specifications
  • Microscope: - Zeiss Axio Observer Z1, fully motorized inverted microscope with an ASI PZ-2000FT XY-Z Piezo stage and Zeiss Definite Focus
  • Objectives: - Zeiss Plan-Apochromat 63x/1.40 oil
    - Zeiss Plan-Apochromat 100x/1.40 oil
  • Epifluorescence light source: - Lumencor Spectra X LED illumination bench
  • Camera: - Hamamatsu ORCA Flash 4.0 V2
  • Confocal system: Complete unit provided by Visitron Systems GmbH, integrating:
    - Yokogawa CSU-W1 confocal head
    - Visitron Systems laser bench (405, 445, 488, 515 and 561 nm)
  • Imaging Software: - Visiview from Visitron Systems GmbH
For live-cell imaging when confocal is not essential or for imaging of fixed samples, we are equipped with a complete epifluorescence inverted microscope integrating an LED illumination bench and taking advantage of the wide field-of-view of the new sCMOS cameras. This is a fully motorized setup that operates with a hardware autofocus system. Flow and temperature control for imaging live fission yeast cells grown in microfluidic chambers is also available on this setup.
    Specifications
  • Microscope: - Zeiss Axio Observer Z1, fully motorized inverted microscope with an ASI PZ-2000FT XY stage and Zeiss Definite Focus.
  • Objectives: - Zeiss Plan-Apochromat 63x/1.40 oil
    - Zeiss Plan-Apochromat 100x/1.40 oil
  • Epifluorescence Light source: - Lumencor Spectra X LED illumination bench
  • Camera: - Hamamatsu ORCA Flash 4.0 V2
  • Imaging Software: - Visiview from Visitron Systems GmbH.
This system is used for routine screening of fission yeast phenotypes on slides or petri dishes using transmitted light.
    Specifications
  • Microscope: - Zeiss Axio Vert A1
  • Objectives: Zeiss A-Plan 10x/0.25 Ph1.
    - Zeiss LD A-Plan 20x/0.35 Ph1.
    - Zeiss A-Plan 40x/0.65 Ph2
We have set up a Linux server to be used with Fiji for standard analysis. It is connected to a dedicated data storage system. Deconvolution using Svi Huygens Essential is available on this platform.
 
Microfluidics
The microfluidic technology that has been implemented in the laboratory allows us to couple high-resolution microscopy with the analysis of single cells grown in an environment that can be modulated in real-time. Using this approach, we can precisely and rapidly alter key parameters such as temperature, medium composition and the concentrations of drugs while monitoring the response of living cells to these inputs.
We have established the entire infrastructure for standard microfabrication in a dedicated room with controlled light, temperature, and limited dust. All the equipment required for developing, prototyping and producing microfluidic devices by soft lithography (including a spincoater, UV insulator, plasma cleaner, high precision hot plates, dedicated microscope, ultrasound water bath, dessicator) is available on the platform. In addition, we are using high-precision pressure controllers and valve matrices to control the flow of medium through the microfluidic chambers as well as a microfluidic-based temperature control system.
Cytometry
For cell cycle analysis and protein detection in populations of cells, we are equipped with a BD Accuri C6 flow cytometer. The cytometer is set up with two lasers (488 and 552 nm) and four detectors (in addition to SSC and FSC). The Accuri is compatible with any type of tube and only requires water as sheath fluid. Analysis of flow cytometry data is performed using Flowjo.
Laboratory Evolution
Laboratory evolution experiments with S. pombe take advantage of a multiplexed system of “ministats” that allows us to maintain up to 32 independent cultures in a controlled environment. This technology has been implemented in the laboratory with the help of Dr. Maitreya Dunham (University of Washington, Seattle, USA). Using this system, cultures can be kept at constant optical density for several months, regardless of their initial generation time. Two incubation blocks have been set up to run parallel experiments at different temperatures. In addition, the various cultures can be maintained in different media. Samples can simply be collected from the effluent line for further analysis, from live-cell imaging to whole genome sequencing.