Center for Advanced Light Microscopy (CALM)
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Spinning disk confocal

UltraView_Vox

Technique

The difference between a point scanning confocal and a spinning disk confocal microscope is the number of image points acquired at the same time. While a point scanning confocal microscope images one spot at a time, a spinning disk confocal microscope illuminates about 1000 spots in parallel and detects emitted light using a camera. The advatage of this approach is an increase in imaging speed and / or reducion of peak power at the illuminated spot. The latter is important for survival of cells during live cell imaging experiments. Imaging many confocal spots at the same time has the disadvantage of loosing sectioning quality. With increasing density of the confocal spots, the probability of photons emitted from a certain focal spot being detected through the pinhole of a nighboring spot rises.

Use

Confocal technique for investigation of fluorescent living cells. Kinetic studies are possible due to the high speed of this technique. Depending on the configuration of the instrument, microirradiation studies for photoactivation / inactivation are possible.

Configuration

Manufacturer: Perkin Elmer
Type: UltraView Vox
Microscope: Zeiss
Spinning Disk Unit: Yokogawa 

Illumination
Lasers, 405 nm, 442 nm, 488 nm, 514 nm, 561 nm, 640 nm
Halogen (DIC/PC)

Optics
all objectives are manufactured by Zeiss
10x 0.25 air objective , Ph1, A-Plan
20x 0.3 air objective
40x 1.3 oil immersion objective, Ph3, DIC, FLUAR
63x 1.4 oil immersion objective DIC, Plan-APOCHROMAT
100x 1.4 oil immersion objective DIC, Plan-APOCHROMAT

Camera
Hamamatsu EMCCD C9100-50 Frame transfer camera
14 bit, 1000 x 1000 eff. Pixel
Pixel Size 8 µm x 8 µm
Int. gain 4.4 e/ADcount, Min. EM-gain 6, Max. 2000
Readout noise 10 e- at min EM gain, less 1 e- at max. EM gain
Dark current 0.0005 e/pixel*s (at -40°C)
Full well capacity 70 000 e-

Image stacksize max z 200 µm