Scanning Confocal Microscope | Department of Biological Sciences

Scanning Confocal Microscope

LSM710 Zeiss Laser Scanning Microscope with Airyscan 1

The LSM710 is equipped with a Zeiss Axio Observer.Z1 inverted microscope and a wide range of lenses including: 5x EC Plan-Neofluar, 20x Plan APO, 10x (water) C-APO, 40x (water) C-APO, and 63x (water) LD C- APO

The illumination and detection design gives users the ability to select up to ten fluorochromes for simultaneous spectral imaging. The 34-channel detector is more sensitive and more flexible than previous detector technologies and yields spectral resolutions down to 3 nm. This spectral detection unit can be adjusted within a wavelength range of 390 to 750 nm. Bandpass windows can be as small as 3.2 nm wide. In Lambda Mode, 34 channels can be simultaneously acquired.

There is a "Smart Setup" option which allows users to select the fluorochromes that they are using from a database of over 600 fluorochromes, to allow for automatic hardware configuration.

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The direct laser coupling "Pigtail" design of up to 8 lasers, allows for minimized laser light loss into the scan head with unique optical geometry in the scan head to improve laser suppression leading to increased sensitivity, improved signal to noise ratio, and easy integration of new lasers.

The configuration of lasers is: a 25mW 405 nm diode laser, a 25 mW multi-line Argon laser (458, 488, and 514 nm), a 20 mW 561 nm DPSS laser, and a 5mW 633 nm HeNe laser.

The spectral recycling loop recovers almost all grating photon losses by reintroducing them into the holographic spectral separation grating which leads to increased sensitivity. The system optics are designed for wavelengths ranging from 350-1100 nm. There is a twin gate main beam splitter. Each wheel has 10 filter positions leading to 100 possible combinations. The movement of scanners across the sawtooth scan field is absolutely linear for equal pixel dwell, therefore no correction is needed.

The minimum resolution is 4 by 1 pixels and the maximum resolution is 6144 by 6144 pixels. The scan can be zoomed from 0.6 times to 40 times in steps of 0.1.

The Airyscan is a special detector module added to the out coupling port of LSM710. It consists of a Gallium Arsenide Phosphid (GaAsp) photomuliplier tube (PMT) array of 32 elements. A resolution gain of 1.7 fold is obtained in all spacial directions.

This system has software, called "ZEN", that can configure all motorized functions and can save and restore application-specific configurations.

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