Fig. 1: Correlative single nucleosome imaging. | Nature Communications

Fig. 1: Correlative single nucleosome imaging.

From: Correlative single molecule lattice light sheet imaging reveals the dynamic relationship between nucleosomes and the local chromatin environment

Fig. 1

A Schematic of lattice light sheet microscopy. B A sample slice of single nucleosomes (middle) and their associated chromatin environment (right) from a 3D volume of cell nucleus (left). Circles indicate tracked nucleosomes. Colors represent the z position relative to the focal plane. Scale bar = 1000 nm C The trajectory of the nucleosome in the blue box in (B). Top: Single nucleosome trajectory tracked across five consecutive 20 ms frames; Bottom: Overlay of nucleosome trajectory on the chromatin channel. D Schematic comparing lattice light sheet microscopy voxel size (dashed cube), nucleosome localization precision (magenta sphere), and the potential nucleosome density (gray disks). E Chromatin density classification. Top left: deconvolved chromatin image; Top right: corresponding chromatin density classification; Bottom left: histogram of chromatin intensity in arbitrary units for top deconvolved image. Blue curve indicates combined Gaussian Mixture Model fitting and red curves indicate individual components; Bottom right: histogram of the proportion of voxels assigned to different chromatin density classes for cell displayed in top image. Scale bar = 1000 nm. Bar height indicates mean and error bars represent standard deviation. Dots represent proportion from a single time point. N = 1 biological replicate taken across 50 consecutive time points. F Representative mean square displacement of nucleosomes in log-log scale for a single cell. The dashed lines show the linear fitting to a power law relationship (\({{{{{\rm{MSD}}}}}}=4D{\Delta {{{{{\rm{t}}}}}}}^{{{{{{\rm{\alpha }}}}}}}\)) where \({{{{{\rm{\alpha }}}}}}\), \({{{{{\rm{D}}}}}}\) and \(\Delta {{{{{\rm{t}}}}}}\) are the anomalous alpha exponent, the apparent diffusion coefficient respectively and the time lag. Colors indicate different chromatin density classes with the same convention as in (E).

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