title: Reconstruction of explicit structural properties at the nanoscale via spectroscopic microscopy.
abstract: The spectrum registered by a reflected-light bright-field spectroscopic microscope (SM) can quantify the microscopically indiscernible, deeply subdiffractional length scales within samples such as biological cells and tissues. Nevertheless, quantification of biological specimens via any optical measures most often reveals ambiguous information about the specific structural properties within the studied samples. Thus, optical quantification remains nonintuitive to users from the diverse fields of technique application. In this work, we demonstrate that the SM signal can be analyzed to reconstruct explicit physical measures of internal structure within label-free, weakly scattering samples: characteristic length scale and the amplitude of spatial refractive-index (RI) fluctuations. We present and validate the reconstruction algorithm via finite-difference time-domain solutions of Maxwell's equations on an example of exponential spatial correlation of RI. We apply the validated algorithm to experimentally measure structural properties within isolated cells from two genetic variants of HT29 colon cancer cell line as well as within a prostate tissue biopsy section. The presented methodology can lead to the development of novel biophotonics techniques that create two-dimensional maps of explicit structural properties within biomaterials: the characteristic size of macromolecular complexes and the variance of local mass density.
authors.affiliation: Northwestern University, Technological Institute, Department of Biomedical Engineering, E310, 2145 Sheridan Road, Evanston, Illinois 60208, United States.
authors.forename: Lusik
authors.initials: L
authors.lastname: Cherkezyan
authors.affiliation: Northwestern University, Technological Institute, Department of Biomedical Engineering, E310, 2145 Sheridan Road, Evanston, Illinois 60208, United States.
authors.forename: Di
authors.initials: D
authors.lastname: Zhang
authors.affiliation: Northwestern University, Technological Institute, Department of Biomedical Engineering, E310, 2145 Sheridan Road, Evanston, Illinois 60208, United States.
authors.forename: Hariharan
authors.initials: H
authors.lastname: Subramanian
authors.affiliation: Northwestern University, Technological Institute, Department of Electrical Engineering, L359, 2145 Sheridan Road, Evanston, Illinois 60208, United States.
authors.forename: Allen
authors.initials: A
authors.lastname: Taflove
authors.affiliation: Northwestern University, Technological Institute, Department of Biomedical Engineering, E310, 2145 Sheridan Road, Evanston, Illinois 60208, United States.
authors.forename: Vadim
authors.initials: V
authors.lastname: Backman
mesh_terms: D000465:Algorithms; D019073:HT29 Cells; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D008297:Male; D008853:Microscopy / Q000379:methods*; D036103:Nanotechnology / Q000379:methods*; D061848:Optical Imaging / Q000379:methods*; D011467:Prostate / Q000737:chemistry; D015203:Reproducibility of Results; D012815:Signal Processing, Computer-Assisted