05. PhaseView light sheet

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 nm
• ET525/50
• ET600/50
• 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 watery refractive index media (volume < 15ml. Agarose strips containing sampes can be mounted in water and observed straight away.

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