Source: http://aoot.osa.org/oe/abstract.cfm?uri=oe-25-13-15428
Timestamp: 2019-04-21 02:13:26+00:00

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We have developed a novel technique to quantify submicron scale mass density fluctuations in weakly disordered heterogeneous optical media using confocal fluorescence microscopy. Our method is based on the numerical evaluation of the light localization properties of an ‘optical lattice’ constructed from the pixel intensity distributions of images obtained with confocal fluorescence microscopy. Here we demonstrate that the technique reveals differences in the mass density fluctuations of the fluorescently labeled molecules between normal and cancer cells, and that it has the potential to quantify the degree of malignancy of cancer cells. Potential applications of the technique to other disease situations or characterizing disordered samples are also discussed.
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Fig. 1 Construction of a disordered optical lattice from confocal imaging (schematic pictures): (a’) Imaging of a sample with a laser beam. (a) Voxel-wise scanning on the xy- plane (z = constant) to construct a confocal 2D plane image of a DAPI stained cell nucleus targeting DNA molecules. (b) A typical confocal image- 2D micrograph. (c) A sample disordered optical lattice of the size of confocal image: each dot in the optical lattice is determined from the pixel intensity values in confocal fluorescence image, as shown in (b).
Fig. 2 Schematic flowchart for comparing the structural disorder using confocal micrographs. (i) The confocal images of the nuclei of two samples were obtained. (ii) Optical lattices are constructed and eigenvalues are obtained by solving the Anderson tight binding model Hamiltonian. (iii) The structural disorder of the samples are then obtained by calculating the inverse participation ratio (IPR) of the systems from the eigenfunctions in a Gaussian color noise model and compared.
Fig. 3 (a), (b), and (c): Representative confocal images of a normal astrocyte, an astrocyte progenitor, and a U87 astrocytoma cell nuclei, respectively. (a’), (b’) and (c’): Their corresponding Lsd(<IPR>) images (2D IPR plot) at sample length L = 0.4 µm (we have taken Lsd(<IPR>) = <IPR>). The scale bar in the confocal image corresponds to 5 µm.
Fig. 4 Bar plots for mean Lsd(<IPR>) values (n = 12-15 cells, 3-5 micrographs per cell, 3 sets) for the normal astrocyte, astrocyte progenitor, and U87 astrocytoma cells nuclei at sample length L = 1.6 µm. Student’s t-test obtained p-value < 0.05 for each pair.
Fig. 5 Structural disorder at different sample length scales (L) (sample size L × L). n = 12-15 cells for each type of normal astrocyte, astrocyte progenitor, and U87 astrocytoma cells, where 3-5 confocal micrographs for each cell around mid-nucleus were considered for the analysis.
Fig. 6 Schematic flowchart for IPR calculation and 2D IPR image construction from confocal fluorescence image.
(2) ε(x,y)= dn(x,y) n 0 ∝ d I CFM (x,y) < I CFM > .
(4) 〈 IPR(L) 〉 L×L = 1 N ∑ i=1 N ∫ 0 L ∫ 0 L E i 4 (x,y)dxdy .
(5) 〈 IPR(L) 〉 ∝ L sd = 〈 d n 2 〉 1/2 × l c .

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