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The vast majority of the microscopy methods currently available to studybiological samples require staining prior to imaging. Nevertheless, the abilityto reveal specific cell structures or organelles in a label-free manner remainsdesirable in different contexts. Polarization microscopy has long beenconsidered as an interesting alternative to fluorescence-based methods in orderto gain specificity on the imaged biological samples. In this work, we show howan original polarization imaging technique, implementing micro-wave photonicsand referred to as orthogonality-breaking (OB) microscopy, can provideinformative polarization images from a single scan of the cell sample in a fastand sensitive way. For OB imaging, the sample is probed with a laser setupsimultaneously generating two orthogonal polarizations shifted in frequency bya few tens of MHz. If the imaged samples display some polarimetric properties,the orthogonality between the two polarizations is broken, leading to abeatnote interference signal that can be detected with a fast detector. Thecomparison of the images of various cell lines at different cell-cycle stagesobtained by OB polarization microscopy and fluorescence confocal images showsthat an endogenous polarimetric contrast arizes on compacted chromosomes duringcell division. This technique paves the way to label-free real-timepolarization imaging of mitotic chromosomes with further potential applicationsin histology and cancer diagnosis.

Label-free microscopy of mitotic chromosomes using the polarization Orthogonality Breaking technique