Point spread function (PSF) engineering-based methods to enhance resolution and contrast of optical microscopes have experienced great achievements in the last decades. These techniques include: stimulated emission depletion (STED), time-gated STED (g-STED), ground-state depletion microscopy (GSD), difference confocal microscopy, fluorescence emission difference microscopy (FED), switching laser mode (SLAM), virtual adaptable aperture system (VAAS), etc. Each affords unique strengths in resolution, contrast, speed and expenses. We explored how PSF engineering generally could be used to break the diffraction limitation, and concluded that the common target of PSF engineeringbased methods is to get a sharper PSF. According to their common or distinctive principles to reshape the PSF, we divided all these methods into three categories, nonlinear PSF engineering, linear PSF engineering, and linear-based nonlinear PSF engineering and expounded these methods in classification. Nonlinear effect and linear subtraction is the core techniques described in this paper from the perspective of PSF reconstruction. By comparison, we emphasized each method’s strengths, weaknesses and biologic applications. In the end, we promote an expectation of prospective developing trend for PSF engineering.

Point spread function (PSF) engineering-based methods to enhance resolution and contrast of optical microscopes have experienced great achievements in the last decades. These techniques include: stimulated emission depletion (STED), time-gated STED (g-STED), ground-state depletion microscopy (GSD), difference confocal microscopy, fluorescence emission difference microscopy (FED), switching laser mode (SLAM), virtual adaptable aperture system (VAAS), etc. Each affords unique strengths in resolution, contrast, speed and expenses. We explored how PSF engineering generally could be used to break the diffraction limitation, and concluded that the common target of PSF engineeringbased methods is to get a sharper PSF. According to their common or distinctive principles to reshape the PSF, we divided all these methods into three categories, nonlinear PSF engineering, linear PSF engineering, and linear-based nonlinear PSF engineering and expounded these methods in classification. Nonlinear effect and linear subtraction is the core techniques described in this paper from the perspective of PSF reconstruction. By comparison, we emphasized each method’s strengths, weaknesses and biologic applications. In the end, we promote an expectation of prospective developing trend for PSF engineering.