<p>Biodynamic imaging (BDI) is a high-content optical imaging technology based on Fourier-domain digital holography and Doppler spectroscopy of intracellular dynamics. There are three main functions of the BDI technique, which are optical coherence imaging (OCI), motility contrast imaging (MCI) and tissue dynamics spectroscopy (TDS). OCI is related to <em>en face</em> optical coherence tomography (OCT) using partially coherent speckle generated by broad-area illumination with coherence detection through digital holography. MCI provides noninvasive functional imaging by treating intracellular motility as an endogenous dynamic imaging contrast agent. TDS produces broad-band Doppler fluctuation power spectra that contain the ensemble of all intracellular motions by collecting and extracting depth-resolved quasi-elastic dynamic light scattering from inside multicellular living tissue. This thesis presents the development and applications of BDI systems. Doppler spectral clustering analysis is demonstrated when comparing fresh canine lymphoma biopsies and their corresponding flash-Frozen samples. Doppler spectral phenotyping analysis is used to identify a non-predictive phenotype of TDS that shows a systemic red-shift of frequencies. Doppler spectral shift analysis is used to monitor bacterial infection of living tissue. </p>

<p>Intracellular motions are important signatures of living tissues, and intracellular dynamics reflect overall cell function and health. Traditional microscopy methods can track 2D cellular motions but do not provide an ensemble evaluation of intracellular activity. Biodynamic imaging (BDI) is a unique 3D imaging technique based on the phase shifts of dynamic light scattering and is highly sensitive to intracellular dynamics in living cells and their changes. This makes BDI a versatile tool to evaluate many different types of samples under various scenarios, and BDI has the potential to improve patient diagnosis and to provide valuable information for health care research. This may include evaluating sample activity, profiling patient chemotherapy response, and studying drug mechanisms. This thesis discusses the theory and modeling of BDI, the construction of BDI systems, sample heterogeneity analysis (TDSI), and the use of BDI to study cytoskeletal drug mechanisms, improve embryo selection and select therapies in pre-clinical trials.</p>

<p>Infertility is the disease of the reproductive system and is estimated to affect more than 10% of the people of reproductive age. Assisted reproductive technologies (ART) are methods designed to alleviate infertility problems. <i>In vitro </i>embryo production is part of most infertility treatments and the efficiency of ART is low due to the lack of reliable methods to measure embryo viability. In order to improve the success rate of ART procedures, the current study was designed to investigate the use of an optical analyzer technology known as the Biodynamic Imaging (BDI) system for viability assessment. BDI is a novel approach that is able to measure intracellular dynamic processes that are directly related to functional events. During a series of experiments, 13 different biomarkers of oocytes and embryos were monitored by the BDI microscope and used for machine learning and evaluation of BDI sensitivity. We monitored cellular mechanisms essential for proper embryo development such as (1) extrusion of first and second polar body (2) energy status and mitochondrial activity, and (3) viability of embryos with different cellular composition. We were able to identify several biomarkers that have the potential to indicate viability: (1) slope, (2) NSD, (3) Knee (4) Floor, and (5) R² could consistently differentiate between oocytes and embryos of different viability. In addition, the BDI microscope could successfully predict the energy status of embryos by identifying 4 biomarkers (Slope, Knee, Floor, and Dy). Finally, a lipidomic analysis was done to evaluate the lipid composition of oocytes with different cytoplasm integrities. This analysis demonstrated that there is a difference in lipid subclasses among oocytes with dark vs. light cytoplasm. The results indicate that the BDI is useful for viability assessment of oocytes and embryos and may be helpful for the improvement of the efficiency of assisted reproductive technologies.</p>

<div>Biological dynamics have been studied by many methods. Fluorescence dynamic microscopy and optical coherence tomography provided fundamental understandings of biological systems. However, their high NA optics only represent local characteristics. Biodynamic imaging (BDI) technique implements a low NA optics and acquires the statistical average of Doppler shifts that occurred by dynamic light scattering with biological dynamic subsystems provided globally averaged dynamic characteristics. </div><div>BDI is used for this study to investigate biomedical applications. The chemotherapy efficacy measurement by BDI demonstrated a good agreement between the Doppler spectral phenotypes and the preclinical outcomes. Also, dynamic responses of microbiomes by chemical stimuli demonstrated featured Doppler characteristics. The bacterial infection of epithelial spheroids showed consistent spectral responses and antibiotic-resistant E. coli infection treatment with a sensitive and resistive antibiotic showed a dramatic contrast. Furthermore, the phase-sensitive characteristics of BDI provided a clue to understanding the characteristics of the random process of biological systems. Levy-like heavy-tailed probability density functions are demonstrated and </div><div>the shape changed by infection will be discussed. </div>