Extended Data Fig. 7: Comparison of pixelated and Zernike pupil based in situ PSF estimation of Tetrapod blinking patterns generated from a phase plate.

(a) Comparison of estimated PSF and pupil from bead data and single molecule blinking patterns. The phase patten generated from a phase plate is more complex than the ones from a deformable mirror, both the pixelated and Zernike pupil based modelling methods can retrieve the pupil function from single molecule blinking data. However, the Zernike-based method requires accurate initial values of the Zernike coefficients, otherwise, it fails to retrieve the accurate pupil. (b, c) Estimated Zernike coefficients from Zernike-based in situ PSF modelling with (b) or without (c) accurate initial values of the Zernike coefficients. (d) Example raw camera frame used for in situ PSF modelling. Scale bar, 1 µm (a), 10 µm (d). ADU, analog to digital unit.