A comprehensive method to study the DNA's association with lamin and chromatin compaction in intact cell nuclei at super resolution

Abstract

We expanded the fBALM (DNA structure fluctuation-assisted binding activated localization microscopy) method by developing a stable methodological sequence that enables dual-color imaging of high-resolution genomic DNA together with LaminB1.

Repair and misrepair of telomeric DNA in dynamic interactions with PML nuclear bodies and lamin B1. A gear-wheel nuclear traffic.

“…Condemine [25] also mentions that PML "tracks can cross the nucleus and were frequently observed filing lamina gaps at the nuclear envelope". As well, our results are consistent with the recent paper applying superresolution microscopy showing that lamin B1 has a dashed character with γH2AX-clusters filling its gaps [35], coinciding with the earlier study by Olins showing the "partial colocalization of Lamin B1 and lamin B receptor (LBR) with the tendency of intermittent dashed pattern" [36,37].…”

Advanced image-free analysis of the nano-organization of chromatin and other biomolecules by Single Molecule Localization Microscopy (SMLM)

“…The image analysis tools using established algorithms, implemented for instance in the open-source program ThunderSTORM [106] , have also become standard for Single Molecule Localization Microscopy (SMLM). The applications presented in [33] suggest their utility also for the cell nucleus research considered here, especially if the results thus obtained are supported by other experimental methods of molecular biology. However, many biophysical studies beyond molecular biology do not require visualization but quantification of parameters characteristic for a biological system, molecular arrangements, or parts of complex molecularly structured system components.…”

“…Methods to extract mesh like structures can be used for SMLM in the form of Voronoi tessellation [29] , [30] and Delaunay triangulation [31] , [32] . For a Voronoi vertices diagram, the 2D plane is divided into tiles around each point, where the edge is defined by the equidistant bisector to the next point (see for instance [33] ). The Delaunay triangulation splits the plane into triangles with SMLM points as vertices (see for instance [34] ).…”

Phase Imaging of Human Cells Using Digital Holographic Microscopy

“…Fluorescent marking, particle tracking, and phase marking are now being combined to track how DNA (Deoxyribonucleic acid) combines to create and rebuild chromosomes within the human nucleus. Examples being, [25][26][27][28][29][30][31][32].…”