Coherent confocal light absorption and scattering spectroscopic (C-CLASS) microscopy, which extends the principles of light scattering spectroscopy to subcellular imaging, can be used to reveal biological structures well beyond the diffraction limit. Here we show that high-resolution C-CLASS microscopy can be used to detect nanoscale changes in chromatin structure. Unlike most methods for chromatin monitoring, C-CLASS microscopy can be used label-free in live cells. Live differentiating hiPSC organoids were measured over the space of sixteen days and characteristic chromatin changes were observed.
Nanoscale changes in the nuclear structure have been shown to play a critical role in genetic and transcriptional alterations and are a hallmark of neoplasia. Genomic processes are regulated by chromatin packing density, thus underlying the significance of understanding the subnuclear structure and its role in the regulation of molecular processes. However, the dynamic and multiscale aspects of these phenomena have remained an open problem. The key reason is the lack of technologies for label-free nanoscale-sensitive measurements in live cells. We have developed confocal light absorption and scattering spectroscopic (CLASS) microscopy for label-free chromatin sensing in live cells.
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