Confocal Laser Scanning Microscope

​The 8th generation Confocal instruments offers ultimate sensitivity and resolving power for high demanding samples. Classical CLSM has some inherent limitations. Attempt to increase the resolution by making the pinhole smaller will necessarily lead to significant decrees in detected light - consequently - the signal-to-noise ratio (SNR) drops. To balance between resolution, SNR, and acquisition speed, Carl Zeiss developed a new approach to laser scanning confocal microscopy named Airyscan.

Super Resolution Confocal with Airyscan:

Airyscan is a detector that draws on the fact that a fluorescence microscope will image a point-like source as an extended Airy disk (Airy pattern). Airyscan solves this conundrum between resolution and light efficiency by imaging the Airy disk onto a concentrically arranged hexagonal detector array. Its detection area consists of 32 single elements, each of which acts like a very small pinhole. The confocal pinhole itself remains open and does not block light – thus all photons of the whole Airy disk are collected (Fig 2 left). The signals from all detector elements are then reassigned to their correct position, producing an image with increased SNR and resolution. Because it capitalizes on the scanning and optical sectioning capabilities of a confocal, Airyscan works with standard samples and standard dyes, and even with your thicker samples such as tissue sections. 

Figure 2. Super-Resolution and FAST Airyscan modules: 

In the additional Fast mode, you profit from yet another advantage of the area detector. The excitation beam is elongated in y and the Airyscan detector, with just one horizontal scanner movement, acquires four lines of image information instead of only one (Fig 2 right). This parallelization delivers a unique combination of high speed, high resolution and high sensitivity.​
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​Figure 3. Drosophila melanogaster neuromuscular junctions stained for Bruchpilot (BRP). Sample courtesy of J. Pielage, Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland ​
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34 Channels Spectral Imaging:

The Zeiss LSM880 Confocal Microscope is equipped for multi-channel fluorescent imaging. The spectral detector together with the software for spectral unmixing makes it possible to image simultaneously multiple different fluorophores, provided that reference spectra for each of the individual fluorophores are available. The spectral detector also can be used for conventional multi-channel fluorescence imaging. The high sensitivity of the detector allows detection of weak fluorescent signals while employing low levels of laser illumination. In samples with high auto-fluorescent (e.g. chlorophyll in plant cells), the spectral imaging and linear unmixing can be used to record the auto-fluorescent spectral characteristics, and remove it later from the image (Fig 4).