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In this thesis, I have shown the potential of single-molecule imaging to study chromatin architecture. The primary challenge was to systematically identify and characterise different building blocks of the chromatin organisation. Some elements were known since long, for example, the nucleosomes at the lowest level and the metaphase X-shaped chromosomes at the highest level of DNA compaction. In between these two extreme levels, a vast spectrum of structures remained mostly unexplored due to resolution limitation of light microscopy and lack of specificity in electron microscopy. Furthermore, an effort to combine and synchronise observations from different studies on chromatin organisation was lacking. This thesis is an attempt to combine old and new evidence to understand various structure-function conundrums of chromatin organisation and understand how chromatin structure affects gene regulation.