Super-resolution microscopy based on parallel detection

Abstract

Image scanning microscopy based on pixel reassignment can improve the confocal resolution limit without losing the image signal-to-noise ratio (SNR) greatly [C. J. R. Sheppard, \Super-resolution in confocal imaging," Optik 80(2) 53-54 (1988). C. B. Müller, E. J€ org, \Image scanning microscopy, \Phys. Rev. Lett. 104(19) 198101 (2010). C. J. R. Sheppard, S. B. Mehta, R. Heintzmann, \Superresolution by image scanning microscopy using pixel reassignment," Opt. Lett. 38(15) 2889-2892 (2013)]. Here, we use a tailor-made optical¯ber and 19 avalanche photodiodes (APDs) as parallel detectors to upgrade our existing confocal microscopy, termed as parallel-detection super-resolution (PDSR) microscopy. In order to obtain the correct shift value, we use the normalized 2D cross correlation to calculate the shifting value of each image. We characterized our system performance by imaging°uorescence beads and applied this system to observing the 3D structure of biological specimen.

Photon-Weight-Reallocated Stimulated Emission Depletion Nanoscopy

“…(6) Calculating new central-seven channel data sets by respectively multiplying the raw data sets with the spatial weight map. The process also included (7) reconstructing the spatial-weight-reassigned STED (SWR-STED) from the new central-seven channels’ data sets and the reassigned STED (R-STED) from the raw seven channels’ data sets using pixel reassignment method − based on parallel detection. Finally, (8) by replacing HDS and S0 with the SWR-STED and the R-STED, respectively, steps 4 and 5 were repeated to obtain the PSW for the STED.…”

Seeing beyond the limit: A guide to choosing the right super-resolution microscopy technique

“…The resolution of an optical system can be defined as the smallest distance between J o u r n a l P r e -p r o o f two points at which the two can still be distinguished as separate entities. The resolution of super resolution techniques varies widely (Fig 4) -from pixel reassignment techniques, which produce around a 1.4x improvement in resolution or an effective resolution of down to 120nm [35,36], to the recently developed MinFlux technique which has a reported resolution of down to 1nm [34]. Techniques, such as STED and SIM inherently allow for improvements in axial resolution, albeit to different extents; however, SMLM applications will generally require adaptations such as deformation of the point spread function (PSF) to provide axial super resolution [37].…”

Nanoscale fluorescence imaging of biological ultrastructure via molecular anchoring and physical expansion

“…A number of other microscopy techniques, such as pixel reassignment and fluctuation-based super-resolution microscopy, can also generate sub-diffraction-limit images at the far-field. Pixel reassignment super-resolution microscopy, for example, can resolve biological samples at ~ 120–150 nm resolution in the lateral dimension and ~ 350–450 nm resolution in the axial dimension [ 7 , 30 ]. In this technique, an array detector rather than a single-point detector is used to capture the fluorescence signals obtained from the laser scan by using single or multiple focal spots.…”