BioDynamic Imaging of living tissue

Ran An, Purdue University

Abstract

BioDynamic Imaging (BDI) is based on Fourier domain short-coherence gated off-axis digital holography. BDI includes optical coherence imaging (OCI), which permits direct wide-field depth-gated imaging by acquiring en face images at a fixed depth inside scattering media without the need to scan; motility contrast imaging (MCI), which uses intra-cellular motility as an endogenous imaging contrast agent, providing label free, non-invasive functional imaging; tissue dynamics spectroscopy (TDS), which uses fluctuation spectroscopy to generate tissue-response spectrograms that track time-resolved changes in intracellular motility in response to perturbations, providing specific fingerprints for different intracellular responses to the specific perturbations; tissue dynamics spectroscopic imaging (TDSI), which applies TDS to localized areas across the entire tissue to image the heterogeneous localized cellular response to specific perturbations. This dissertation describes the principles and the latest updates of BioDynamic Imaging. It also covers the detailed experimental protocol as well as detailed data processing protocols. An important application of TDSI—deep tissue label-free mitosis detection—is demonstrated. Furthermore, BDI is applied to perform tissue viability assessment of multicellular tumor spheroids using motility contrast imaging. The viability of porcine oocytes was also studies using BDI. Cumulus-oocyte complexes (CoC) were studied, showing the difference between standard empirical assessment and the assessment provided by BDI. The viability of bare oocytes was studied, and principal component analysis was used to distinguish fertilized and unfertilized oocyte groups. The technique was able to distinguish normal unfertilized oocytes from heat shocked (unhealthy) unfertilized oocytes. Interesting results of an oocyte cleavage spectrogram and blastocyst dynamic speckles are shown. In the final part, BDI was applied to doxorubicin chemotherapy assessment of the canine non-Hodgkin lymphoma, and clinical results and BDI results are compared.

Degree

Ph.D.

Advisors

Nolte, Purdue University.

Subject Area

Optics

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