Fig. 1: lIllustration of PAR-SIM setup and synchronization procedure.
a The system rendering and illustration based on the commercial system. b The 2D-SIM excitation setup is Airy Polar-SIM mounted on the Nikon Ti2 ECLIPSE fluorescent microscopy, the diffracted light from SLM translates to back pupil of objective with a transform lens (Tube in excitation), pizza polarization rotator, 0 order stop mask, and a pair of 4f-relay lens (omitted). The pattern excited fluorescence from sample is collected by objective and passes through a multiband dichroic mirror. The detection tube lens images the fluorescent sample right on its focal plane, and the ROI is cut with the custom-designed sub-ROI slit. L1 and L2 are a pair of Fourier relay lens whose magnification is 1.5 (f2:f1 = 1.5). It transforms the selected sub-ROI on the sCMOS camera. The xy-scan galvo mirror set is precisely situated on the middle conjugation plane of L1 and L2. Sub-ROI slit is a custom-designed transparent rectangular window. Obj, objective; DM, dichroic mirror; SLM, spatial light modulator; L1&L2, relay lens1, and lens2; AOn, analog output with n ports giving the analog signals to these controlled optoelectronic devices. c The top half part of sensor should follow the blue orbit to scan the sub-ROI and the bottom should follow the red arrow order to scan, since it is determined by the readout direction of sensor. d The sCMOS opens the half top of the sensor, and the slope readout time coming from the first several patterned sub-ROIs can be parallelly reused as the rest patterned sub-ROIs’ exposure time on these opening up sensor rows. 1st Angle 1st Phase to 1st Angle 3rd Phase (P1-P3, labeled with 1 dark −2 middle-3 light blue rectangle); 2nd Angle 1st Phase to 2nd Angle 3rd Phase (P4-P6, labeled with 4 dark −5 middle-6 light yellow rectangle); 3rd angle 1st phase to 3rd angle 3rd phase (P7-P9, labeled with 7 dark −8 middle-9 light pink rectangle). e The synchronization signal of PAR-SIM. \({{\boldsymbol{T}}}_{{\bf{i}}}\), Galvo stopping time; \({{\boldsymbol{T}}}_{{\bf{ii}}}\), SLM duration; \({{\boldsymbol{T}}}_{{\bf{iii}}}\), SLM high voltage duration; \({{\boldsymbol{T}}}_{{\bf{D}}{\bf{1}}}\), the camera exposure delay signal between the Kth frame and the K + 1th frame; \({{\boldsymbol{T}}}_{{\bf{D}}{\bf{2}}}\), the delay between start timepoint of camera frame exposure and scanner beginning to deflect sub-ROIs; \({{\boldsymbol{T}}}_{{\bf{D}}{\bf{3}}}\), the delay time between the SLM starting and the scanner device starting on each step for buffering the Galvo settling time. In Frame K + 2, the signal schematic diagram is stretched for clearly showing these denoted signal names and corresponding relations. The various colors with numbers correspond to d P1-P9 sub-ROIs
