Single shot approach for three-dimensional imaging with double-helix point spread functions

2016 ◽  
Author(s):  
René Berlich ◽  
Andreas Bräuer ◽  
Sjoerd Stallinga
Keyword(s):  
Double Helix ◽  
Three Dimensional ◽  
Single Shot ◽  
Point Spread Functions ◽  
Three Dimensional Imaging ◽  
Point Spread

scholarly journals Analyzing engineered point spread functions using phasor-based single-molecule localization microscopy

2020 ◽  
Author(s):  
Koen J.A. Martens ◽  
Abbas Jabermoradi ◽  
Suyeon Yang ◽  
Johannes Hohlbein
Keyword(s):  
Saddle Point ◽  
Single Molecule ◽  
Software Package ◽  
Double Helix ◽  
Three Dimensional ◽  
Fourier Plane ◽  
Point Spread Functions ◽  
Localization Microscopy ◽  
Point Spread ◽  
Single Molecule Localization Microscopy

The point spread function (PSF) of single molecule emitters can be engineered in the Fourier plane to encode three-dimensional localization information, creating double-helix, saddle-point or tetra-pod PSFs. Here, we describe and assess adaptations of the phasor-based single-molecule localization microscopy (pSMLM) algorithm to localize single molecules using these PSFs with sub-pixel accuracy. For double-helix, pSMLM identifies the two individual lobes and uses their relative rotation for obtaining z-resolved localizations, while for saddle-point or tetra-pod, a novel phasor-based deconvolution approach is used. The pSMLM software package delivers similar precision and recall rates to the best-in-class software package (SMAP) at signal-to-noise ratios typical for organic fluorophores. pSMLM substantially improves the localization rate by a factor of 2 - 4x on a standard CPU, with 1-1.5·104 (double-helix) or 2.5·105 (saddle-point/tetra-pod) localizations/second.

scholarly journals Single shot three-dimensional imaging using an engineered point spread function

2016 ◽  
Vol 24 (6) ◽  
pp. 5946 ◽  
Author(s):  
René Berlich ◽  
Andreas Bräuer ◽  
Sjoerd Stallinga
Keyword(s):  
Point Spread Function ◽  
Three Dimensional ◽  
Single Shot ◽  
Three Dimensional Imaging ◽  
Point Spread ◽  
Spread Function

Single-shot three-dimensional imaging with a scattering layer [Invited]

2021 ◽  
Vol 60 (10) ◽  
pp. B32
Author(s):  
Guowei Li ◽  
Wanqing Yang ◽  
Yaoming Bian ◽  
Haichao Wang ◽  
Guohai Situ
Keyword(s):  
Three Dimensional ◽  
Single Shot ◽  
Scattering Layer ◽  
Three Dimensional Imaging

scholarly journals Single-shot three-dimensional imaging of dilute atomic clouds

2014 ◽  
Vol 39 (18) ◽  
pp. 5317 ◽  
Author(s):  
Kaspar Sakmann ◽  
Mark Kasevich
Keyword(s):  
Three Dimensional ◽  
Single Shot ◽  
Three Dimensional Imaging

scholarly journals Three-Dimensional Super-Resolution in Eukaryotic Cells Using the Double-Helix Point Spread Function

2017 ◽  
Vol 112 (7) ◽  
pp. 1444-1454 ◽  
Author(s):  
Alexander R. Carr ◽  
Aleks Ponjavic ◽  
Srinjan Basu ◽  
James McColl ◽  
Ana Mafalda Santos ◽  
...  
Keyword(s):  
Point Spread Function ◽  
Double Helix ◽  
Three Dimensional ◽  
Super Resolution ◽  
Eukaryotic Cells ◽  
Point Spread ◽  
Spread Function

scholarly journals Virtual phase conjugation based optical tomography for single-shot three-dimensional imaging

2018 ◽  
Vol 26 (4) ◽  
pp. 3779 ◽  
Author(s):  
Yuta Goto ◽  
Atsushi Okamoto ◽  
Atsushi Shibukawa ◽  
Kazuhisa Ogawa ◽  
Akihisa Tomita
Keyword(s):  
Phase Conjugation ◽  
Optical Tomography ◽  
Three Dimensional ◽  
Single Shot ◽  
Three Dimensional Imaging

scholarly journals Correlations of three-dimensional motion of chromosomal loci in yeast revealed by the double-helix point spread function microscope

2014 ◽  
Vol 25 (22) ◽  
pp. 3619-3629 ◽  
Author(s):  
Mikael P. Backlund ◽  
Ryan Joyner ◽  
Karsten Weis ◽  
W. E. Moerner
Keyword(s):  
Point Spread Function ◽  
Double Helix ◽  
Dynamical Behavior ◽  
Three Dimensional ◽  
Mean Square Displacement ◽  
Single Particle Tracking ◽  
3D Tracking ◽  
Point Spread ◽  
Spread Function ◽  
Diploid Cells

Single-particle tracking has been applied to study chromatin motion in live cells, revealing a wealth of dynamical behavior of the genomic material once believed to be relatively static throughout most of the cell cycle. Here we used the dual-color three-dimensional (3D) double-helix point spread function microscope to study the correlations of movement between two fluorescently labeled gene loci on either the same or different budding yeast chromosomes. We performed fast (10 Hz) 3D tracking of the two copies of the GAL locus in diploid cells in both activating and repressive conditions. As controls, we tracked pairs of loci along the same chromosome at various separations, as well as transcriptionally orthogonal genes on different chromosomes. We found that under repressive conditions, the GAL loci exhibited significantly higher velocity cross-correlations than they did under activating conditions. This relative increase has potentially important biological implications, as it might suggest coupling via shared silencing factors or association with decoupled machinery upon activation. We also found that on the time scale studied (∼0.1–30 s), the loci moved with significantly higher subdiffusive mean square displacement exponents than previously reported, which has implications for the application of polymer theory to chromatin motion in eukaryotes.

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