Quantitative phase imaging: advances to 3D imaging and applications to neuroscience

Kim, Tae Woo

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https://hdl.handle.net/2142/88067 Copy

Description

Title

Quantitative phase imaging: advances to 3D imaging and applications to neuroscience

Author(s)

Kim, Tae Woo

Issue Date

2015-07-17

Director of Research (if dissertation) or Advisor (if thesis)

Popescu, Gabriel

Doctoral Committee Chair(s)

Popescu, Gabriel

Committee Member(s)

• Goddard, Lynford L.
• Boppart, Stephen A.
• Gillette, Martha U.

Department of Study

Electrical & Computer Eng

Discipline

Electrical & Computer Engr

Degree Granting Institution

University of Illinois at Urbana-Champaign

Degree Name

Ph.D.

Degree Level

Dissertation

Keyword(s)

• Microscopy
• Quantitative phase imaging
• Scattering theory
• Interferometry
• Coherence
• Neuroscience
• Cell biology
• Tomography

Abstract

This thesis provides a brief overview of quantitative phase imaging (QPI) methods along with applications and advances made on them. First, spatial light interference microscopy (SLIM) is introduced as a QPI method extensively used in this thesis. Using this setup, an application of QPI in neuroscience is demonstrated by studying the emergent formation of a neuronal network. Second, an expansion of this QPI method into a 3D quantitative imaging method, called white-light diffraction tomography (WDT), has been shown. Lastly, an initial result for another advance in SLIM is introduced by combining SLIM with a programmable illumination. In the first part of this work, the emergent self-organization of a neuronal network has been demonstrated using the SLIM system. The emergent self-organization of a neuronal network in a developing nervous system is the result of a remarkably orchestrated process involving a multitude of chemical, mechanical and electrical signals. Little is known about the dynamic behavior of a developing network (especially in a human model) primarily due to a lack of practical and non-invasive methods to measure and quantify the process. Using the SLIM system, several fundamental properties of neuronal networks have been measured non-invasively from the sub-cellular to the cell population level. This method quantifies network formation in human stem cell derived neurons and shows correlations between trends in the growth, transport, and spatial organization of such a system, by utilizing the quantitative phase data with novel analysis tools, including dispersion-relation phase spectroscopy (DPS). A deeper understanding of neuronal network formation has been provided by studying filopodia dynamics in neurons. By measuring the dry mass change over time and several other new metrics, it is shown that the filopodia dynamics successfully reflect the expected neurite outgrowth. In the second part, white-light diffraction tomography (WDT) is introduced as a new approach for imaging microscopic transparent objects such as live unlabeled cells in 3D. The approach extends diffraction tomography to white light illumination and imaging rather than scattering plane measurements. The experiments were performed using the SLIM system. The axial dimension of the object was reconstructed by scanning the focus through the object and acquiring a stack of phase-resolved images. The 3D structures of live, unlabeled red blood cells are imaged and compared with confocal and scanning electron microscopy images. The 350 nm transverse and 900 nm axial resolution achieved allows us to reveal sub-cellular structures at high resolution in E. coli cells and HT29 cells. Furthermore, a 4D imaging capability, with the fourth dimension being time, has also been demonstrated. The WDT theory is further extended to explain light scattering through thick tissue, which is not in the single scattering regime. The obtained inverse scattering solution for thick samples is then related to the time-reversal theory, and it is proven that there are strong constraints for time-reversal to work. By introducing a few specific examples, including scattering through a particle and scattering through a grating, the physics of light scattering and time-reversal theory is deeply understood. Lastly, an upgrade to the SLIM system with a programmable illumination source, a projector, has been demonstrated. By replacing the ring illumination of PC with a ring-shaped pattern projected onto the condenser plane, results comparable to those of the original SLIM were recovered. This new method minimized the halo artifact of the imaging system by minimizing the effect of spatial coherence caused by the thickness of the illumination. Further application of this technique into optogenetics is introduced and the initial results are presented.

Graduation Semester

2015-8

Type of Resource

text

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http://hdl.handle.net/2142/88067

Copyright and License Information

Copyright 2015 Tae Woo Kim

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