LSM900 Airyscan
- Ki Hng
This microscope is equipped with an Airyscan 2 unit, a 32 channel array detector from Zeiss which can acquire images with up to 8x improvement in Signal to Noise ratio and 1.7x higher resolution than a conventional confocal. It also has 2x multiplex. For more information on how it works, please refer to Zeiss product details
To use the Airyscan, you can either use Smart Setup, selecting Airyscan, and selecting either Resolution, Speed or Sensitivity mode, or by adding Airyscan channels in "Imaging Setup"
Zoom needs to be set to at least 1.8. The pinhole automatically adjusts to twice the airy unit in order for all the light to go to the scanner instead of being eliminated by the pinhole.
Prior to imaging in SR mode, the light from the sample must be automatically centered on the detector before acquisition. To open the Airyscan Detector Adjustment toolbar (which was under Maintain in Zen Black on the Multiphoton), click on the Airyscan icon at the bottom of the screen. Tick ‘Adjust in live and continuous scans’ and then start a continuous scan so the Airyscan centers and the icon goes green, and then untick it before moving the sample to where you actually want to image. it is best to do this at 512 x 512, with speed at maximum - on a bright sample.
This allows the the scanner to centre the airy signal on the 32 detectors. Once this is centred and you can see the bright spot in the center of the 32 detectors as above, uncheck the 'Adjust in Continuous Scan" box. This is to avoid the airyscan continuously recentering unsuccessfully when you scan less bright samples.
You can now change the frame size and scan speed.
When setting up your imaging settings, it is advised to image at 16-bit, and to only use 25% of the dynamic range. Using too much of the dynamic range increases processing artefacts when your images are put through Airyscan processing.
Airyscan SR and Airyscan SR-2Y data has to be Airyscan processed to get the final image. Airyscan processing can be done on the same computer, or on Marvin,