University of Heidelberg
BIOQUANT

ViroQuant-CellNetworks RNAi Screening Facility

Microscopy

 

"ready for DAQ" assays → high-throughput microscopy

 

Screening microscopes are used for high-throughput / high-content data acquisition. The facility provides three Olympus IX81 (Scan^R / Cell^R) and two Leica TCS SP5 microscopes. The microscopes are equipped with standard as well as screening data acquisition software.

 

Olympus IX81

High-throughput screening

 

Wide field microscopes
  • 3x   Olympus Biosystems IX81 'inverted'

 

Wide field microscopes enable very rapid scanning procedures as well as time series of biological assays. For high-throughput wide field screening the microscopes can be loaded with various well plate formats.

- Two setups are equipped with incubator chambers allowing temperature, humidity and CO2 regulation for long term in vivo measurements.

- One setup facilitates automated plate loading via a stacker/gripper system for up to 40 well plates.

Leica TCS SP5
Perkin Elmer Opera LX

High-content screening

 

Confocal laser scanning microscopes
  • 2x   Leica TCS SP5 DMI6000 'inverted'
Spinning disk confocal screening microscope
  • Perkin Elmer Opera LX 'spinning disk'

 

In order to increase the information content, optical sectioning is indespensable. Confocal setups are used for high-content confocal screening providing 3-dimensional data stacks as well as time series of biological assays. The two Leica systems are equipped with an incubator chamber allowing temperature, humidity and CO2 regulation for long term in vivo measurements.

 

 

 

Microscopes

Wide field screening microscopes

 

Olympus 1 (room 312, wall)   Olympus Biosystems IX81 'inverted'

  • illumination: Mercury (Hg) + Xenon (Xe) lamp
  • detection: wide field
  • sensor: CCD
  • channels: 6 (DAPI, CFP, GFP, Cy3, Cy5, DAPI/FITC/TxRed)
  • software: Scan^R / xcellence (data acquisition and analysis); versions: 2.3.0.5 (Scan^R data acquisition) | 1.3.0.3 (Scan^R analysis)
  • probe handling: Object slides, standardised MTPs, cell arrays and adapter for various plate formats
  • environment control: none
  • automated plate loading: stacker/gripper system (up to 40 well plates)
  • data storage:  access to BioQuant internal mass storage (Large Scale Data Facility - LSDF)

 

Olympus 2 (room 312, door)   Olympus Biosystems IX81 'inverted'

  • illumination: Mercury (Hg) + Xenon (Xe) lamp
  • detection: wide field
  • sensor: CCD
  • channels: 6 (DAPI, CFP, GFP, Cy3, Cy5, DAPI/FITC/TxRed)
  • Software: Scan^R / xcellence (data acquisition and analysis); versions: 2.3.0.5 (Scan^R data acquisition) | 1.3.0.3 (Scan^R analysis)
  • probe handling: Object slides, standardised MTPs, cell arrays and adapter for various plate formats
  • environment control: incubator chamber allowing temperature, humidity and CO2 regulation
  • automated plate loading: none
  • data storage:  access to BioQuant internal mass storage (Large Scale Data Facility - LSDF)

 

Olympus 3 (room 222, S2 area)   Olympus Biosystems IX81 'inverted'

  • illumination: Mercury (Hg) + Xenon (Xe) lamp
  • detection: wide field
  • sensor: CCD
  • channels: 5 (DAPI, YFP, Cy5, CFP, GFP)
  • Software: Scan^R / xcellence (data acquisition and analysis); versions: 2.3.0.5 (Scan^R data acquisition) | 1.3.0.3 (Scan^R analysis)
  • probe handling: Object slides, standardised MTPs, cell arrays and adapter for various plate formats
  • environment control: incubator chamber allowing temperature, humidity and CO2 regulation
  • automated plate loading: none
  • data storage:  access to BioQuant internal mass storage (Large Scale Data Facility - LSDF)

Confocal microscope

 

SP5 (room 222, S2 area)   Leica TCS SP5 DMI6000 'inverted'

  • illumination: monochromatic light covering UV and visible spectrum:

         - Diode laser (50 mW @ 405 nm)

         - Ar+ laser (100 mW @ 458, 476, 488, 496, 514 nm)

         - HeNe laser (1-10 mW @ 543, 594, 633 nm)

         - DPSS laser (10 mW @ 561 nm)

  • detection: confocal (continuously variable pinhole)
  • sensor: photomultiplier (PMT)
  • channels: whole VIS via Acousto Optical Beam Splitter (AOBS) and prism spectral detector (max. 5 channels in parallel)
  • scanner: switch between conventional mode (5 fps @ 512x512 resolution) and resonant mode (25 fps @ 512x512)
  • Software: Leica LAS AF (data acquisition and analysis, version 1.8.2)
  • probe handling: Object slides, standardised MTPs, cell arrays and adapter for various plate formats
  • environment control: incubator chamber allowing temperature, humidity and CO2 regulation
  • data storage:  access to BioQuant internal mass storage (Large Scale Data Facility - LSDF)

Confocal screening microscope

 

SP5 (room 312)   Experimental Leica TCS SP5 DMI6000 'inverted'

  • illumination: monochromatic light covering UV and visible spectrum:

         - Diode laser (50 mW @ 405 nm)

         - Ar+ laser (100 mW @ 458, 476, 488, 496, 514 nm)

         - HeNe laser (1-10 mW @ 543, 594, 633 nm)

         - DPSS laser (10 mW @ 561 nm)

  • detection: confocal (continuously variable pinhole)
  • sensor: photomultiplier (PMT)
  • channels: whole VIS via Acousto Optical Beam Splitter (AOBS) and prism spectral detector (max. 5 channels in parallel)
  • scanner: switch between conventional mode (5 fps @ 512x512 resolution) and resonant mode (25 fps @ 512x512)
  • Software: Leica LAS AF (data acquisition and analysis) & Matrix Screener for automated confocal screening
  • probe handling: Object slides, standardised MTPs, cell arrays and adapter for various plate formats.
  • environment control:  incubator chamber allowing temperature, humidity and CO2 regulation
  • data storage:  access to BioQuant internal mass storage (Large Scale Data Facility - LSDF)

 

 

Spinning disk confocal screening microscope

 

Opera LX (room 222, S2 area)   Perkin Elmer Opera LX 'spinning disk'

  • illumination: monochromatic light covering visible spectrum:

         - Diode laser (75 mW @ 488 nm)

         - Diode laser (40 mW @ 561 nm)

         - Diode laser (40 mW @ 640 nm)

  • detection: confocal point scanning (spinning disk)
  • objective lenses: 20x (NA=0.7), 60x (NA=1.2). Water immersion, automated water supply
  • sensor: CCD camera (1.3 MegaPixel, 12bit)
  • channels: whole VIS via dichroit / triple notch filter
  • Software: Acapella. Online image processing
  • probe handling: Object slides, standardised MTPs, cell arrays and adapter for various plate formats.
  • data storage:  access to BioQuant internal mass storage (Large Scale Data Facility - LSDF)

 

 

Contact: E-Mail (Last update: 27/06/2013)