Contribution of advanced fluorescence nano microscopy towards revealing mitotic chromosome structure

Abstract

The organization of chromatin into higher-order structures and its condensation process represent one of the key challenges in structural biology. This is important for elucidating several disease states. To address this long-standing problem, development of advanced imaging methods has played an essential role in providing understanding into mitotic chromosome structure and compaction. Amongst these are two fast evolving fluorescence imaging technologies, specifically fluorescence lifetime imaging (FLIM) and super-resolution microscopy (SRM). FLIM in particular has been lacking in the application of chromosome research while SRM has been successfully applied although not widely. Both these techniques are capable of providing fluorescence imaging with nanometer information. SRM or “nanoscopy” is capable of generating images of DNA with less than 50 nm resolution while FLIM when coupled with energy transfer may provide less than 20 nm information. Here, we discuss the advantages and limitations of both methods followed by their contribution to mitotic chromosome studies. Furthermore, we highlight the future prospects of how advancements in new technologies can contribute in the field of chromosome science.

The Toolbox for Untangling Chromosome Architecture in Immune Cells

“…Fluorescence and electron imaging techniques are evolving rapidly for elucidating chromatin organization. There are excellent reviews on this topic, which will not be discussed in detail here (40,41,70,71). Although these cutting-edge imaging techniques have not been widely applied to studying the immune system, with the improvement of resolution and throughput, they will be sure to illuminate the connection between genome organization and immune cell activity.…”