Probing biology on its home turf: Tools for ethology and protein dynamics
Ravan, Aniket S.
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Permalink
https://hdl.handle.net/2142/122143
Description
Title
Probing biology on its home turf: Tools for ethology and protein dynamics
Author(s)
Ravan, Aniket S.
Issue Date
2023-12-01
Director of Research (if dissertation) or Advisor (if thesis)
Gruebele, Martin
Chemla, Yann
Doctoral Committee Chair(s)
Gruebele, Martin
Chemla, Yann
Committee Member(s)
Shukla, Diwakar
Nelson, Mark
Department of Study
School of Molecular & Cell Bio
Discipline
Biophysics & Quant Biology
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Keyword(s)
Animal behavior
Protein dynamics
Computer Vision
Machine Learning
Microscopy
Abstract
Biological systems exist out of equilibrium by constantly exchanging information and energy with their environment. This exchange is characterized by complex interactions at multiple length and time scales. Top-down approaches seeking a mechanistic understanding of biological processes require quantitative observations in the full context of these interactions. This requirement motivates the need to engineer tools that enable probing biological systems closer to their native environment. This work includes efforts in developing two such tools that allow probing the larval zebrafish system in the context of biophysical questions.
The thesis is broadly divided into three parts, one part for the introduction, and one part each focusing on one of the techniques. In Chapter 1, I will motivate the need to probe biological systems close to their native environment. Chapter 1 forms the first part of the thesis. Chapter 2-3, I will discuss a novel technique to perform behavioral experiments and pose estimation of larval zebrafish in a 3-D cubic tank. A convolutional neural network model predicts larval poses across various experiments rapidly using a digitally generated and broadly applicable training dataset. In Chapter 4, I will perform objective behavioral analysis of 3-D larval behaviors and compare them with behaviors observed in classical 2-D locomotion assays. The 3-D behavioral analysis reveals the ability of the larva to feint when startled. Chapters 2-4 form the second part of the thesis. In Chapter 5, using the larval zebrafish system, I will broadly describe a technique to perform temperature-dependent FRET measurements of proteins in single cells of live zebrafish on an epifluorescence microscope, allowing observations of protein-protein interactions and changes in protein stability.
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