Faculty of Science

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Light Sheet Microscopy

The General Instrumentation Department hosts the following light sheet microscope.

PhaseView Alpha3 (room HG01.228)

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Microscope:

  • Manual upright Olympus BX43F microscope

Excitation: Dual light sheet illumination through two 10x/0.25NA long distance Olympus objectives using a 4-laser combiner of the following laser lines:

  • 405nm/50mW
  • 488nm/50mW
  • 561nm/50mW
  • 638nm/100mW

Emission Selection: The system has the following filters mounted on a motorized filter wheel:

  • ET440/40
  • ET525/50
  • ET600/50m
  • ET690/50
  • Notch filter 405/488/561/638

Detection: ORCA-FLASH4.0V3

  • Scientific CMOS Sensor 2048 (H)×2048 (V), 6.5 μm×6.5 μm
  • QE 82 % (@560 nm)
  • Format 13.312 mm×13.312 mm
  • Camera Link interface (100fps)

Objectives:

  • 10x/0.3 Water dipping long distance
  • 20x/0.5 Water dipping long distance
  • 40x/0.8 Water dipping long distance

Chamber and Sample holders: Chamber dimensions: width 21mm, length 70m height 25mm. Sample size up to 10 x 10 x 10 mm, cuvette filled with medium (volume < 15ml.  Chamber highly resistant to corrosive medium: embryo medium, saltwater, clearing medium

Motorized Rotation Module: Motorized rotation module with 1024 steps resolution, software controlled.

XY Tiling: XY motorized stage with 15mm range and 0.1um precision. Software controlled

Software: QtSPIM software for image acquisition XY, Z, channels, t (time), θ (rotation) and tiling

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