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From Molecules to Organs : Microscopy and Multi-Scale Nature of Development

Citation

Trivedi, Vikas (2015) From Molecules to Organs : Microscopy and Multi-Scale Nature of Development. Dissertation (Ph.D.), California Institute of Technology. doi:10.7907/Z9SX6B59. https://resolver.caltech.edu/CaltechTHESIS:05292015-080950114

Abstract

Morphogenesis is a phenomenon of intricate balance and dynamic interplay between processes occurring at a wide range of scales (spatial, temporal and energetic). During development, a variety of physical mechanisms are employed by tissues to simultaneously pattern, move, and differentiate based on information exchange between constituent cells, perhaps more than at any other time during an organism's life. To fully understand such events, a combined theoretical and experimental framework is required to assist in deciphering the correlations at both structural and functional levels at scales that include the intracellular and tissue levels as well as organs and organ systems. Microscopy, especially diffraction-limited light microscopy, has emerged as a central tool to capture the spatio-temporal context of life processes. Imaging has the unique advantage of watching biological events as they unfold over time at single-cell resolution in the intact animal. In this work I present a range of problems in morphogenesis, each unique in its requirements for novel quantitative imaging both in terms of the technique and analysis. Understanding the molecular basis for a developmental process involves investigating how genes and their products- mRNA and proteins-function in the context of a cell. Structural information holds the key to insights into mechanisms and imaging fixed specimens paves the first step towards deciphering gene function. The work presented in this thesis starts with the demonstration that the fluorescent signal from the challenging environment of whole-mount imaging, obtained by in situ hybridization chain reaction (HCR), scales linearly with the number of copies of target mRNA to provide quantitative sub-cellular mapping of mRNA expression within intact vertebrate embryos. The work then progresses to address aspects of imaging live embryonic development in a number of species. While processes such as avian cartilage growth require high spatial resolution and lower time resolution, dynamic events during zebrafish somitogenesis require higher time resolution to capture the protein localization as the somites mature. The requirements on imaging are even more stringent in case of the embryonic zebrafish heart that beats with a frequency of ~ 2-2.5 Hz, thereby requiring very fast imaging techniques based on two-photon light sheet microscope to capture its dynamics. In each of the hitherto-mentioned cases, ranging from the level of molecules to organs, an imaging framework is developed, both in terms of technique and analysis to allow quantitative assessment of the process in vivo. Overall the work presented in this thesis combines new quantitative tools with novel microscopy for the precise understanding of processes in embryonic development.

Item Type:Thesis (Dissertation (Ph.D.))
Subject Keywords:Developmental biology, 2-photon light sheet, Hybridization Chain Reaction, Cardiac development, Cartilage development
Degree Grantor:California Institute of Technology
Division:Biology and Biological Engineering
Major Option:Bioengineering
Thesis Availability:Public (worldwide access)
Research Advisor(s):
  • Fraser, Scott E. (co-advisor)
  • Pierce, Niles A. (co-advisor)
Thesis Committee:
  • Ravichandran, Guruswami (chair)
  • Gharib, Morteza
  • Phillips, Robert B.
  • Fraser, Scott E.
  • Pierce, Niles A.
Defense Date:19 May 2015
Non-Caltech Author Email:viktri.iitk (AT) gmail.com
Record Number:CaltechTHESIS:05292015-080950114
Persistent URL:https://resolver.caltech.edu/CaltechTHESIS:05292015-080950114
DOI:10.7907/Z9SX6B59
Related URLs:
URLURL TypeDescription
http://dx.doi.org/ncomms7798DOIArticle adapted for ch. 4
http://dx.doi.org/10.1364/BOE.6.002056DOIArticle adapted for ch. 7
http://dx.doi.org/10.1016/j.bpj.2012.11.1140DOIArticle adapted for ch. 8
ORCID:
AuthorORCID
Trivedi, Vikas0000-0003-0953-0553
Default Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:8917
Collection:CaltechTHESIS
Deposited By: Vikas Trivedi
Deposited On:19 Jan 2016 22:00
Last Modified:04 Oct 2019 00:08

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