Localization Precision of Three-Dimensional Superresolution Fluorescence Imaging Using a Double-Helix Point Spread Function

2009 ◽  
Author(s):  
Matthew Lew ◽  
Michael A. Thompson ◽  
Majid Badieirostami ◽  
W. E. Moerner
Keyword(s):  
Fluorescence Imaging ◽  
Point Spread Function ◽  
Double Helix ◽  
Three Dimensional ◽  
Point Spread ◽  
Spread Function ◽  
Localization Precision

scholarly journals Three-dimensional localization precision of the double-helix point spread function versus astigmatism and biplane

2010 ◽  
Vol 97 (16) ◽  
pp. 161103 ◽  
Author(s):  
Majid Badieirostami ◽  
Matthew D. Lew ◽  
Michael A. Thompson ◽  
W. E. Moerner
Keyword(s):  
Point Spread Function ◽  
Function Versus ◽  
Double Helix ◽  
Three Dimensional ◽  
Point Spread ◽  
Spread Function ◽  
Localization Precision

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 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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