Confocal microscopes work to block out of focus light by utilising the pinhole and therefore generate clearer images than those from a widefield system. Laser scanning confocal microscopes (LSCM) utilise a detector and point scanning system. Functional imaging is available on the Leica SP5, and multiphoton imaging on the Zeiss 980. Airyscan and Airyscan 2 detectors on the Zeiss confocals allow for increased resolution.
The Zeiss 880 LSCM microscope.
Inverted microscope
Environmental control - temperature, humidity & CO2
Airyscan detector
405 nm
458 nm
488 nm
514 nm
543 nm
594 nm
633 nm
Airyscan technology delivers superresolution with high sensitivity (4-8x) at 140 nm laterally and 400 nm axially resolution (1.7x) whilst preserving precious emission light normally rejected at a closed pinhole. This gentle superresolution imaging with increased sensitivity and speed reduces phytotoxicity and bleaching and allows long-term live cell imaging without artefacts caused by influencing the sample’s viability and biological function.
Zeiss 980 LSCM - Airyscan 2
Zeiss 880 LSCM - Airyscan
Zeiss 800 LSCM - Airyscan
The Airyscan detector draws on the fact that a fluorescence microscope will image a point-like source of light as an extended Airy disk or Airy pattern.
In a standard confocal microscope the out-of-focus emission light is rejected at the pinhole. You get a sharper image by closing the pinhole to reject out-of-focus light, however, it’s also much dimmer as a great deal of light is then lost. The smaller the pinhole, the higher the resolution, but the greater the loss in light. Normally on a confocal a compromise is required.
The Airyscan solves this trade off between resolution and light efficiency by imaging the entire Airy disk onto a concentrically-arranged hexagonal detector array consisting of 32 single detector elements, all of which act like very small pinholes. The signals from all the detector elements are then reassigned to their correct position, producing an image with increased signal-to-noise ratio and super-resolution.
Airyscan imaging capitalises on the scanning and optical sectioning capabilities of a confocal and therefore works with standard samples, standard dyes and importantly even with thicker samples such as tissue sections or whole animal mounts that need a higher penetration depth.
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