Center for Advanced Light Microscopy (CALM)
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Light sheet

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Technique

Light sheet microscopy differs from other microscopic techniqus by the ortogonal orientation of the illumination relative to the detection beam path. By using optical tricks, the excitation can be confined to one plane. In front and behind this plane no excitation takes place. The emitted light from this plane is imaged by the detection optics. This type of microscopy is inherently confocal and provides the best preservation of the specimen because photobleaching and phototoxicity is reduced to a minimum. Some implementations of light scheet microscopy facilitate the investigation of the sample from different view angles. By using reconstruction algorithms, a 3D representation of the sample with isotropic resolution can be gernerated.

Use

Long term imaging of living organisms, imaging of larger 3D volumes

Configuration

Manufacturer: Homemade (OpenSpim with some modifications)

Excitation: 488 nm and 561 nm Laser

Light sheet generation: Cylinder lens and 10x NA 0.3 water immersion objective  (UMPLFLN 10XW, Olympus)

Detection: 20x NA 0.5 water immersion objective ( (UMPLFLN 20XW, Olympus) and Orac Flash 4.0 V2 Camera

Software: Micro-Manager / Fiji