After the Elyra PS.1 super-resolution, the I2MC Imaging facility has acquired a new high-resolution module Airyscan implemented on an LSM 780 (Zeiss) confocal microscope.
In addition to its two standard PMT detectors, this microscope is equipped with a high-sensitivity GaAsP-type detector. Thanks to its quantum efficiency, which is twice as effective as that of a standard PMT, this microscope is capable of acquiring images from low light intensities. Building on this sensitivity, the installation of the Airyscan enables the acquisition of high-resolution images (140 nm laterally (xy) compared to 200 nm in confocal mode).
The Airyscan detector is particularly useful when the sample, due to weak and/or labile labelling, cannot be imaged in super-resolution (risk of photobleaching). Finally, the ability to switch from confocal to Airyscan mode on the same sample (fixed or living) makes it easier to use.
Illustrations
Microtubule network (15nm) in RPE1 cells: imaged by LSM780 confocal microscopy (left), Airyscan high resolution (centre) and SIM-Elyra PS.1 super-resolution (right). Scale bar 10µm. ©Déborah MICHEL, I2MC
Microtubules (15nm) on RPE1 cells: by confocal microscopy LSM780 (left) and super-resolution SIM-Elyra PS.1 (right). ©Déborah MICHEL, I2MC
As a reminder, super-resolution microscopy techniques
- Structured Illumination Microscopy (SIM): xy resolution = 90nm; z = 350nm / conventional fluorophores
- Photoactivated Localisation Microscopy (PALM) : xy resolution = 20 nm; z = 50 nm / photoactivatable organic probes (eGFP tag, LifeActin)
- direct Stochastic Reconstruction Microscopy (dSTORM): xy resolution = 20 nm; z = 50 nm / standard fluorophores (AF647), specialised buffers, custom-built setup
