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double-helix point spread function microscope  (Double Helix)

 
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    Structured Review

    Double Helix double-helix point spread function microscope
    Double Helix Point Spread Function Microscope, supplied by Double Helix, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/double-helix+point+spread+function+microscope/double+helix+point+spread+function+microscope/pm40239646-639-13-13
    Average 90 stars, based on 1 article reviews
    double-helix point spread function microscope - by Bioz Stars, 2026-09
    90/100 stars

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    Related Articles

    Microscopy:

    Article Title: Live-cell three-dimensional single-molecule tracking reveals modulation of enhancer dynamics by NuRD.
    Article Snippet: .. Analysis of the 20 ms exposure tracks of single CHD4 molecules using our approach revealed a fast unconfined state that was freely Nature Structural & Molecular Biology | Volume 30 | November 2023 | 1628–1639 1630 Article https://doi.org/10.1038/s41594-023-01095-4 b A ng le = Z p os iti on Double-helix point spread function microscopy Sample trajectories Confined Unconfined 0 μm –2 μm +2 μm 10 μm Holo-NuRD complex Separate NuRD subcomplexes Remodeler CHD4 HDAC subcomplex HDAC MBD3 GATA D2A/2B MTA p46/ p48 CHD4 a + MBD3 – MBD3 Step 1 Compute four biophysical parameters using a sliding window for a given trajectory Step 3 GMM classification into two classes: Confined and Unconfined Step 4 Classify subtrajectories by the posterior probability P Sliding window α Fr eq ue nc y Confined Unconfined Unsegmented trajectory Segmented trajectory X(10) 5 100 X(0) Time (s) Po st er io r p ro ba bi lit y P 1.0 0.8 0.6 0.4 0.2 0 Confined Unconfined OutputInput Step 2 Compile biophysical parameters from all sliding windows for all trajectories Trajectory 1 Trajectory 2 Trajectory 3 Trajectory N Dapp (μm 2 s–1) norm||V|| (μm)Lc (nm) α Dapp (μm 2 s–1) norm||V|| (μm)Lc (nm) Dapp (μm 2 s–1) norm||V|| (μm)Lc (nm) 0.7 0.8 0.9 1.0 1.1 2 4 6 8 10 α Time (s) 2 4 6 8 10 Time (s) 2 4 6 8 10 Time (s) 2 4 6 8 10 Time (s) 0.02 0.04 0.06 0.08 0.1 0.2 0.3 0.4 0.5 40 60 80 100 120 140 160 c 0.4 0.8 1.2 120 ||V|| (μU, ∑U) (μC, ∑C) Lc α D 80 40 0 0 0.4 0.8 1.2 120 80 40 160 0.5 1.5 2.5 100 0 300 200 3.5 2 4 6 8 100 500 300 100 700 0 100 300 200 0.5 1.5 2.5 3.5 200 4000 500 300 100 700 2 4 6 8 100 500 300 100 700 500 300 100 0 700 200 400 Fr eq ue nc y Fig. 1 | Live-cell imaging to study NuRD complex binding kinetics and function. a, Schematic representation of the NuRD complex interacting with chromatin in the presence and absence of MBD3. b, Left, single JF549-HaloTagged molecules in the NuRD complex were tracked in 3D using a double-helix point spread function microscope; two puncta are recorded for each fluorophore with their midpoint providing the lateral x, y position and the angle between them representing the axial position in z relative to the nominal focal plane (see Extended Data Fig. 2 and Supplementary Videos 1–8 for examples of the raw data). ..

    Article Title: Live-cell 3D single-molecule tracking reveals how NuRD modulates enhancer dynamics
    Article Snippet: .. We used a double-helix point spread function microscope to generate 3D tracks of single NuRD-HaloTag-JF 549 complexes through a 4 μm slice of the nucleus, so that we could reliably determine biophysical parameters ( ). (Tracking in 2D reduces the length of trajectories because molecules can move in/out of the imaging plane during the experiment.) ..

    Article Title: High-resolution dynamic imaging of chromatin DNA communication using Oligo-LiveFISH.
    Article Snippet: In brief Oligo-LiveFISH is an approach that can track diverse genomic loci at high spatial (20 nm) and temporal (50 ms) resolution and can work in diverse cell types, including primary cells.. Combined with super-localization microscopy and dynamic modeling, Oligo-LiveFISH reveals distinct modes of chromatin communication and the dynamic relationship between transcription and enhancer-promoter interactions.

    Article Title: Live-cell three-dimensional single-molecule tracking reveals modulation of enhancer dynamics by NuRD
    Article Snippet: .. We used a double-helix point spread function microscope to record 3D tracks of single NuRD-HaloTag-JF 549 complexes as they moved through a 4-μm slice of the nucleus (Fig. ) at two distinct temporal regimes: 20 ms and 500 ms (Extended Data Fig. ). ..

    Article Title: Effect of loops on the mean-square displacement of Rouse-model chromatin
    Article Snippet: .. [19] M. P. Backlund, R. Joyner, K. Weis, and W. E. Moerner, Correlations of three-dimensional motion of chromosomal loci in yeast revealed by the double-helix point spread function microscope, Mol. ..

    Article Title: Chromosome Dynamics in Response to DNA Damage.
    Article Snippet: Recent advances in both the technologies used to measure chromatin movement and the biophysical analysis used to model them have yielded a fuller understanding of chromatin dynamics and the polymer structure that underlies it.. Changes in nucleosome packing, checkpoint kinase activation, the cell cycle, chromosomal tethers, and external forces acting on nuclei in response to external and internal stimuli can alter the basal mobility of DNA in interphase nuclei of yeast or mammalian cells.. Although chromatin movement is assumed to be necessary for many DNA-based processes, including gene activation by distal enhancer–promoter interaction or sequence-based homology searches during double-strand break repair, experimental evidence supporting an essential role in these activities is sparse.

    Article Title: Autobiography of W. E. (William Esco) Moerner.
    Article Snippet: .. Mikael P. Backlund, Ryan Joyner, Karsten Weis, and W. E. Moerner, “Correlations of three-dimensional motion of chromosomal loci in yeast revealed by the double-helix point spread function microscope,” Mol. ..

    Live Cell Imaging:

    Article Title: Live-cell three-dimensional single-molecule tracking reveals modulation of enhancer dynamics by NuRD.
    Article Snippet: .. Analysis of the 20 ms exposure tracks of single CHD4 molecules using our approach revealed a fast unconfined state that was freely Nature Structural & Molecular Biology | Volume 30 | November 2023 | 1628–1639 1630 Article https://doi.org/10.1038/s41594-023-01095-4 b A ng le = Z p os iti on Double-helix point spread function microscopy Sample trajectories Confined Unconfined 0 μm –2 μm +2 μm 10 μm Holo-NuRD complex Separate NuRD subcomplexes Remodeler CHD4 HDAC subcomplex HDAC MBD3 GATA D2A/2B MTA p46/ p48 CHD4 a + MBD3 – MBD3 Step 1 Compute four biophysical parameters using a sliding window for a given trajectory Step 3 GMM classification into two classes: Confined and Unconfined Step 4 Classify subtrajectories by the posterior probability P Sliding window α Fr eq ue nc y Confined Unconfined Unsegmented trajectory Segmented trajectory X(10) 5 100 X(0) Time (s) Po st er io r p ro ba bi lit y P 1.0 0.8 0.6 0.4 0.2 0 Confined Unconfined OutputInput Step 2 Compile biophysical parameters from all sliding windows for all trajectories Trajectory 1 Trajectory 2 Trajectory 3 Trajectory N Dapp (μm 2 s–1) norm||V|| (μm)Lc (nm) α Dapp (μm 2 s–1) norm||V|| (μm)Lc (nm) Dapp (μm 2 s–1) norm||V|| (μm)Lc (nm) 0.7 0.8 0.9 1.0 1.1 2 4 6 8 10 α Time (s) 2 4 6 8 10 Time (s) 2 4 6 8 10 Time (s) 2 4 6 8 10 Time (s) 0.02 0.04 0.06 0.08 0.1 0.2 0.3 0.4 0.5 40 60 80 100 120 140 160 c 0.4 0.8 1.2 120 ||V|| (μU, ∑U) (μC, ∑C) Lc α D 80 40 0 0 0.4 0.8 1.2 120 80 40 160 0.5 1.5 2.5 100 0 300 200 3.5 2 4 6 8 100 500 300 100 700 0 100 300 200 0.5 1.5 2.5 3.5 200 4000 500 300 100 700 2 4 6 8 100 500 300 100 700 500 300 100 0 700 200 400 Fr eq ue nc y Fig. 1 | Live-cell imaging to study NuRD complex binding kinetics and function. a, Schematic representation of the NuRD complex interacting with chromatin in the presence and absence of MBD3. b, Left, single JF549-HaloTagged molecules in the NuRD complex were tracked in 3D using a double-helix point spread function microscope; two puncta are recorded for each fluorophore with their midpoint providing the lateral x, y position and the angle between them representing the axial position in z relative to the nominal focal plane (see Extended Data Fig. 2 and Supplementary Videos 1–8 for examples of the raw data). ..

    Binding Assay:

    Article Title: Live-cell three-dimensional single-molecule tracking reveals modulation of enhancer dynamics by NuRD.
    Article Snippet: .. Analysis of the 20 ms exposure tracks of single CHD4 molecules using our approach revealed a fast unconfined state that was freely Nature Structural & Molecular Biology | Volume 30 | November 2023 | 1628–1639 1630 Article https://doi.org/10.1038/s41594-023-01095-4 b A ng le = Z p os iti on Double-helix point spread function microscopy Sample trajectories Confined Unconfined 0 μm –2 μm +2 μm 10 μm Holo-NuRD complex Separate NuRD subcomplexes Remodeler CHD4 HDAC subcomplex HDAC MBD3 GATA D2A/2B MTA p46/ p48 CHD4 a + MBD3 – MBD3 Step 1 Compute four biophysical parameters using a sliding window for a given trajectory Step 3 GMM classification into two classes: Confined and Unconfined Step 4 Classify subtrajectories by the posterior probability P Sliding window α Fr eq ue nc y Confined Unconfined Unsegmented trajectory Segmented trajectory X(10) 5 100 X(0) Time (s) Po st er io r p ro ba bi lit y P 1.0 0.8 0.6 0.4 0.2 0 Confined Unconfined OutputInput Step 2 Compile biophysical parameters from all sliding windows for all trajectories Trajectory 1 Trajectory 2 Trajectory 3 Trajectory N Dapp (μm 2 s–1) norm||V|| (μm)Lc (nm) α Dapp (μm 2 s–1) norm||V|| (μm)Lc (nm) Dapp (μm 2 s–1) norm||V|| (μm)Lc (nm) 0.7 0.8 0.9 1.0 1.1 2 4 6 8 10 α Time (s) 2 4 6 8 10 Time (s) 2 4 6 8 10 Time (s) 2 4 6 8 10 Time (s) 0.02 0.04 0.06 0.08 0.1 0.2 0.3 0.4 0.5 40 60 80 100 120 140 160 c 0.4 0.8 1.2 120 ||V|| (μU, ∑U) (μC, ∑C) Lc α D 80 40 0 0 0.4 0.8 1.2 120 80 40 160 0.5 1.5 2.5 100 0 300 200 3.5 2 4 6 8 100 500 300 100 700 0 100 300 200 0.5 1.5 2.5 3.5 200 4000 500 300 100 700 2 4 6 8 100 500 300 100 700 500 300 100 0 700 200 400 Fr eq ue nc y Fig. 1 | Live-cell imaging to study NuRD complex binding kinetics and function. a, Schematic representation of the NuRD complex interacting with chromatin in the presence and absence of MBD3. b, Left, single JF549-HaloTagged molecules in the NuRD complex were tracked in 3D using a double-helix point spread function microscope; two puncta are recorded for each fluorophore with their midpoint providing the lateral x, y position and the angle between them representing the axial position in z relative to the nominal focal plane (see Extended Data Fig. 2 and Supplementary Videos 1–8 for examples of the raw data). ..

    Imaging:

    Article Title: Live-cell 3D single-molecule tracking reveals how NuRD modulates enhancer dynamics
    Article Snippet: .. We used a double-helix point spread function microscope to generate 3D tracks of single NuRD-HaloTag-JF 549 complexes through a 4 μm slice of the nucleus, so that we could reliably determine biophysical parameters ( ). (Tracking in 2D reduces the length of trajectories because molecules can move in/out of the imaging plane during the experiment.) ..

    other:

    Article Title: Live-cell three-dimensional single-molecule tracking reveals modulation of enhancer dynamics by NuRD.
    Article Snippet: We used a double-helix point spread function microscope25 to record 3D tracks of single NuRD-HaloTag-JF549 complexes as they moved through a 4-μm slice of the nucleus (Fig. 1b) at two distinct temporal regimes: 20 ms and 500 ms (Extended Data Fig. 2).



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