Circular intensity differential scattering of light for biophotonics applications

This thesis demonstrate the application of a label-free, non-invasive biophysical method based on angle-resolved light scattering calculations to characterize different biological samples; virus particles, chromatin fiber, and hierarchical chiral polymers. Chapter 1 provides an introduction on the foundation of electromagnetic theory, light scattering phenomenon, Mueller scattering matrix and its applications to characterize various samples based on numerical simulations and experimental measurements. A numerical method to calculate light scattering quantities, discrete dipole approximation (DDA) method is discussed. In this thesis we have performed the electromagnetic scattering calculations using the DDA method implemented as ADDA code. Chapter 2 demonstrates the angle-resolved circularly polarized light scattering calculations to characterize virus model particles. A coronavirus particle is modeled as having a spherical shaped envelope with cylindrical spikes projected from the envelope surface, and the single-stranded RNA genome polymer has been mimicked with a toroidal helix. The influence of genome polymer packaged as a standard helix in the virion core is also demonstrated. We investigated four different electromagnetic models: (i) a nucleated sphere with spikes that is a coronavirus particle, (ii) a nucleated sphere with no spikes, (iii) a homogeneous sphere, and (iv) a respiratory fluid containing a virus particle. The angular pattern of scattered circularly polarized light, the circular intensity differential scattering of light (CIDS), served as a particle's signature. This scattering signature is found sensitive to the chiral parameters that reveal information about the particles. The effect of changes in the RNA polymer, changes in its packaging, number of turns, handedness, and size are demonstrated on the scattering calculations. Additionally, the extinction efficiency, the depolarization ratio, the total scattered intensity, and the effect of changes in the wavelength of incident light on these scattering quantities are investigated. This biophysical method can offer a label-free identification of virus particles and can help understand their interaction with light. Chapter 3 focus on the the characterization of chromatin organization. Understanding the structural organization of chromatin is essential to comprehend the gene functions. The chromatin organization changes in the cell cycle, and it conforms to various compaction levels. We investigated a chromatin solenoid model with nucleosomes shaped as cylindrical units arranged in a helical array. The solenoid with spherical-shaped nucleosomes was also modeled. The changes in chiral structural parameters of solenoid induced different compaction levels of chromatin fiber. We calculated the angle-resolved scattering of circularly polarized light to probe the changes in the organization of chromatin fiber in response to the changes in its chiral parameters. The electromagnetic scattering calculations were performed using discrete dipole approximation (DDA). In the chromatin structure, nucleosomes have internal interactions that affect chromatin compaction. The merit of performing computations with DDA is that it takes into account the internal interactions. We demonstrated sensitivity of the scattering signal's angular behavior to the changes in these chiral parameters: pitch, radius, the handedness of solenoid, number of solenoid turns, the orientation of solenoid, the orientation of nucleosomes, number of nucleosomes, and shape of nucleosomes. These scattering calculations can potentially benefit applying a label-free polarized-light-based approach to characterize chromatin DNA and chiral polymers at the nanoscale level. Chapter 4 demonstrates the differential scattering of circularly polarized light to characterize the macromolecular structures consisting of hierarchical chirality. We modeled the B-DNA structure composed of a double-helix and a base-pairs helical structure. The angle-resolved scattering of circularly polarized light calculated for the B-DNA shows the additive behavior of the scattering signal contributed from the two individual chirality levels of B-DNA structure, a double-helix and a base-pairs helix. This additive behavior of angle-resolved scattering signal has also been demonstrated for other macromolecular structures comprising different chirality levels; a biological cell is also mimicked as a nucleated sphere, a sphere with a helical nucleus in its core. The individual chiral features of a structure add up to the angle-resolved scattering signal of circularly polarized light produced by the parent structure. These electromagnetic wave scattering calculations can offer a label-free approach to characterize chiral macromolecular structures with hierarchical chirality.