Approach to multiparticle parallel tracking in thick samples with three-dimensional nanoresolution

Abstract

This Letter proposes a method referred to as distorted grating (DG) and double-helix point spread function (DH-PSF) combination microscopy (DDCM), which is capable of multiparticle parallel localization and tracking in a transparent sample thicker than 10 μm, the thickness of cells. A special phase mask, combining the field depth extension capabilities of DG with the three-dimensional (3D) nanolocalization capabilities of the DH-PSF, is designed for multiparticle parallel localization. Time-lapse tracking of one particle moving along the z axis and parallel tracking of two particles are simulated. Results demonstrate that, with only a single snapshot, particles can be localized, tracking with 3D nanoresolution wherever they are. The theoretical localization precisions of DDCM, DH-PSF, and multifocus microscopy are compared. DDCM results in almost constant localization precisions in all three dimensions for a depth of field larger than 10 μm. DDCM is expected to become a tool in investigations of important dynamic events in living cells.

Single shot, three-dimensional fluorescence microscopy with a spatially rotating point spread function

“…The performance of the PSFengineered system is mostly dominated by the adopted CGH and the aberration in the optical system. Since Greengard et al [20] promoted the rotating PSF in 2006, the double-helix PSF has enjoyed great attention in multiple imaging technologies [24,25] due to its high performance in depth estimation. The mainstream CGHs to generate the double-helix PSFs in these applications are generally derived from Gaussian-Laguerre (GL) modes, which were proved to be energy-efficient [26].…”

Recent progress on single-molecule localization microscopy

“…2B ). This method shows the depth of field up to 2 µm, which is higher than astigmatism and the bi-plane approach (Backlund et al 2013 ; Chen et al 2013 ; Grover et al 2011 , 2012 ; Thompson et al 2010 ). Recently, another PSF engineering method called Tetrapod was reported and the depth of field could be extended to several micrometers without large deterioration of localization precision (Gustavsson et al 2018 ; Shechtman et al 2015 , 2017 ).…”

Localization-based full-field microscopy: how to attain super-resolved images

“…The localization based microscopy attempts to extract image information by sampling target fluorescence with super-localized light fields that are much smaller than the diffraction limit and reconstructing the sampled data with localization-specific deconvolution algorithms. The possibility of subdiffraction limited resolution was shown using metamaterials 5 , through statistical anaylsis 6 , with an sCMOS camera 7 , and for particle localization in 3D 8 9 10 . Various superlenses have been proposed in the related context and developed for various applications 11 : for example, by way of the coupling between evanescent diffracted wave and propagating Bloch wave of photonics crystals 12 and by super-oscillation of a band-limited wave induced by diffraction grating 13 .…”