Abstract
Polarized light scanning microscopy is a non-invasive and contrast-enhancing technique to investigate anisotropic specimens and chiral organizations. However, such arrangements suffer from insensitivity to confined blend of structures at sub-diffraction level. Here for the first time, we present that the pixel-by-pixel polarization modulation converted to an image phasor approach issues an insightful view of cells to distinguish anomalous subcellular organizations. To this target, we propose an innovative robust way for identifying changes in the chromatin compaction and distortion of nucleus morphology induced by the activation of the lamin-A gene from Hutchinson–Gilford progeria syndrome that induces a strong polarization response. The phasor mapping is evaluated based on the modulation and phase image acquired from a scanning microscope compared to a confocal fluorescence modality of normal cell opposed to the progeria. The method is validated by characterizing polarization response of starch crystalline granules. Additionally, we show that the conversion of the polarization-resolved images into the phasor could further utilized for segmenting specific structures presenting various optical properties under the polarized light. In summary, image phasor analysis offers a distinctly sensitive fast and easy representation of the polarimetric contrast that can pave the way for remote diagnosis of pathological tissues in real-time.
Highlights
The perception of chromatin compartments as chiral-group structures composed of DNA double helix and proteins inside eukaryotic nuclei is crucial to identify enriched expressed genes regions based on different compaction levels of c hromatin[1,2]
Chromatin compaction level is fascinating to investigate through polarization that is sensitive to the molecular organization, cellular/nuclear shape and very interesting if we want to diagnose genetic pathology such as progeria syndrome as a progressive genetic disorder
The label free polarization technique suffers from information spatial averaging that is insensitive to different mixture of structures in the confined illumination volume defined by Point Spread Function (PSF)
Summary
The perception of chromatin compartments as chiral-group structures composed of DNA double helix and proteins inside eukaryotic nuclei is crucial to identify enriched expressed genes regions based on different compaction levels of c hromatin[1,2]. As there is no known cure, people with progeria due to complications of severe atherosclerosis, either cardiac disease or stroke, typically live to an age of mid-teens to early twenties[4,5,6] It has been an interesting bio sample for image microscopy as a suitable study case to show how a deformation in the cell structures induced by a bad transcription of the genes. In case of cells label-free imaging, it becomes arduous to identify different cell types or/and identify cell structures like the chromatin in nuclear membrane To address this problem, we denote that any modulated signal sequence can be analyzed in frequency domain by Fourier Transform mapped into a single point on a phasor plot. We propose a graphical highly sensitive analytical method for polarimetric images in order to recognize the discrimination at different stages of development in cells
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