Motion-resistant structured illumination microscopy based on principal component analysis.

ABSTRACT

Structured illumination microscopy (SIM) has become one of the most significant super-resolution techniques in bioscience for observing live-cell dynamics, thanks to fast full-field imaging and low photodamage. However, artifact-free SIM super-resolution reconstruction requires precise knowledge about variable environment-sensitive illumination parameters. Conventional algorithms typically, under the premise of known and reliable constant phase shifts, compensate for residual parameters, which will be easily broken by motion factors such as environment and medium perturbations, and sample offsets. In this Letter, we propose a robust motion-resistant SIM algorithm based on principal component analysis (mrPCA-SIM), which can efficiently compensate for nonuniform pixel shifts and phase errors in each raw illumination image. Experiments demonstrate that mrPCA-SIM achieves more robust imaging quality in complex, unstable conditions compared with conventional methods, promising a more compatible and flexible imaging tool for live cells.