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Krieger: BioPipeline High Content Imaging

krieger biopipelline setup

Summary

Krieger is an imaging setup dedicated for live cell imaging. The BioPipeline is fully automated and includes an incubator that can store 44 multi-well plates and a robotic system that automatically mounts the plate on the microscope stage. The microscope is equipped for the image modalities: brightfield, differential interference contrast (DIC), widefield fluorescence imaging and spinning disk confocal imaging. For example, your samples can be stored in the incubator for weeks and can be monitored daily with your preferred imaging modality. We create an automated acquisition pipeline (JOBS) for you and train you how to use the JOB independently.

Image modalities

  • Brightfield
  • DIC
  • Widefield fluorescence imaging
  • Spinning disk confocal imaging

Applications

  • Prolonged fluorescence live cell imaging. There are 6 different laser lines available for the widefield fluorescence and spinning disk confocal mode. A fluorescence mode can also be combined with brightfield or DIC.
  • Highthrough put screening / high-content screening. For live or fixed samples in multi-well plates with any of the imaging modes.
  • Organoid development imaging. Organoids can be monitored using any of the imaging modes (or a combination of imaging modes).
  • Live cell drug screening. Effects of the drugs can be monitored with any of the imaging modes (or a combination of imaging modes).

Technical description

 

Specifications

Microscope body

 Eclipse Ti2-E

Stage

 Automated

Perfect Focus system (PFS)

 Yes

Incubator

 Okolab cage incubator (37⁰C, CO 2)

 Robotic plate mounter

 44 multi-well plate incubator

Objectives

Correction, Magnification, Numerical Aperture, Working Distance

 Plan Apo lambda 4x NA 0.20 air WD 20000µm

 Plan Apo lambda 10x NA 0.45 air WD 4000µm

 Plan Apo lambda 20x NA 0.75 air WD 1000µm

 *Apo LWD Lambda S 20XC WI NA 0.95 WD 950µm

 Plan Apo lambda 40x NA 0.95 air WD 250µm

 *Plan Apo VC 60x WI NA 1.2 WD 310µm

 * automated water dispenser installed

 DIC compatible with 10x, 20x, 40x, and 60x magnifications

Software

 NIS-Elements software (version 5.21.01, High content analysis package)

Lasers

 7 line solid state laser light engine: 405nm, 446nm, 477nm, 520nm, 546nm, 638nm, 749nm (Lumencor Celesta)

Spinning Disk

 X-Light V3 50 µm pinholes (CrestOptics). Can be removed for widefield mode

Emission filters

 Quad dichroic (for laser lines 405 nm, 477 nm, 546 nm and 638 nm)

 440/40nm, 520/20nm, 607/34nm, 694/35nm,809/81nm

 Dual dichroic (for laser lines 446 nm and 520 nm).

 464/23nm, 547/30nm

 Individual filters

 450/40nm, 485/20nm, 520/40nm, 560/25nm, 590/50nm, 700/75nm, 800/81nm

Detector

 ORCA-Fusion digital CMOS camera (Hamamtsu)

 Pixel Size 6.5 μm× 6.5 μm

 Effective number of pixels 2304× 2304

 Effective Area 14.976 mm× 14.976 mm

 Quantum efficiency

 @ 400 nm 65%

 @ 550 nm 80%

 @ 700 nm 70%

 @ 800 nm 50%

DIC/Brightfield

 Diascopic LED

Location

Leichtag building, Room 470