This system is a high-content imaging system providing both spinning disk confocal and widefield imaging. It is optimized for imaging multi-well plates and has both air and water-immersion objectives. It also has temperature and CO2 for live-cell imaging.

Objectives:

Installed

• Plan Apo Lambda 4x/0.2NA (not currently installed)
• Plan Apo 10x/0.45NA
• Apo LWD 20x/0.95NA WI  (0.99-0.9 WD)
• Apo LWD 40x/1.15NA WI (0.61 – 0.59 WD)

Not installed:

• Plan Apo Lambda 2x/0.1NA

4 positions available, WI objectives requires service engineer to exchange and calibrate.

Equipment Details:

• ImageXpress Confocal HT. ai High-Content Imaging System
• Andor Zyla 4.5 camera
• Dual disk unit allowing selection between 3 modes A) 60 micron pinhole disk for your basic confocal requirements B) 50 micron slit for high throughput confocal requirements and C) wide field (not-confocal) imaging mode.
• Gas Mixer
• MetaXpress Imaging and Analysis software

Light Source:

89 North LDI laser illumination

Excitation Lasers:

• 405
• 445
• 475
• 520
• 555
• 640
• 730

Emission Filters:

• DAPI
  • Excitation: 405/20 nm
    Emission: 452/45 nm
    Dichroic: FF409-Di03
• CFP
  • Excitation: 445/20 nm
    Emission: 483/32 nm
    Dichroic: ZT 445/520
• FITC
  • Excitation: 467.5/21 nm
    Emission: 520/28 nm
    Dichroic: ZT 405/470/555/640/730 or FF506-Di03
• YFP
  • Excitation: 520/10 nm
    Emission: 562/40 nm
    Dichroic: ZT 445/520
• TRITC
  • Excitation: 555 nm
    Emission: 598/25 nm
    Dichroic: ZT 405/470/555/640/730
• TxRed
  • Excitation: 555 nm
    Emission: 624/40 nm
    Dichroic: FF593-Di03-25
• Cy5
  • Excitation: 638/17 nm
    Emission: 692/40 nm
    Dichroic: ZT 405/470/555/640/730
• Cy7
  • Excitation: 725/30 nm
    Emission: 794/32 nm
    Dichroic: ZT 405/470/555/640/730

Interface Requirements

Firmware

PC

Software

Reccomendations

To know when you get started

• Knowing your plate dimensions is critical! If you have specific information about your plate it will make the autofocusing easier and smoother
• An empty plate is also important for measuing the dimensions of the plate for autofocusing

Plates known to work well

• Corning 96-well PDL coat (https://ecatalog.corning.com/life-sciences/b2c/US/en/Microplates/Assay-Microplates/BioCoat-Microplates/Corning%C2%AE-BioCoat%C2%AE-Poly-D-Lysine-Microplates/p/354640) reference number 354640
  • Used by the Kampmann lab for growing and imaging Neurons

Resources