Center for Advanced Light Microscopy (CALM)
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Light sheet

slider light sheet 762_735x250

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

Luxendo InVi SPIM


Laser:
488nm direct modulated
561nm direct modulated

Objective:
Dectection: Nikon CFI Apo 25x W 1.1 NA water immersion objective
Illumination: Nikon CFI Plan Fluor 10x W 0.3 NA water immersion objective

Magnification changer:
1.25x, resulting in 31.3x total magnification
2.5x resulting in 62.5x total magnification

Light sheet thickness:
Changeable between 2 – 6 µm

Camera 1:
Hamamatsu Orca Flash 4.0 V3
2048 x 2048 pixels of 6.5 µm x 6.5 µm size
maximum frame rate >80 frames/sec

Camera 2:
For second detection channel
Data see Camera 1

Environmental control:
Ibidi gas mixer for control of CO2 and humidity
Peltier-based temperature control (range 20-37°C)

Computer:
128 GB RAM
Intel Dual Six Core CPU
RAID controller for data streaming, 8 x 4 TB local storage in RAID 0
Nvidia GeForce GTX 1080 graphics card