In an attempt to increase the amount of light passing through the pinhole (0.6AU) to the detector, the laser power was increased. The time series on the left hand side was acquired using the standard LSM (GaAsP detector) with the pinhole closed to 0.6AU. The laser power was increased to 8x that of the Airyscan detector to improve the SNR sufficiently to enable a good deconvolution. The image on the right hand side is the Airyscan time series. The consquence of increasing the laser power in the left hand image is obvious; extremely bad bleaching artefacts.
Sample:U2OS Cells with Lifeactin; Max projection of 6 plane z-stack, 50 volumes over 17.5 minutes Airyscan is clearly the gentle imaging choice