Development of an Automated Optofluidic Microscopy System and its Applications in Reproductive and Developmental Biology

An optofluidic microscopy platform has been developed to perform experiments in reproductive and developmental biology. This platform consists of two separate but related microscopy systems, Robolase I and Robolase IV. Specifically, this platform allows for the following. 1. Custom microfluidic chambers can be used to manipulate sperm and egg independently with automated computer controlled flow rates and valve timings (Robolase I or Robolase IV; Chapters 4-7). 2. Optical scissors can be used to damage individual organelles within an oocyte, egg, or embryo (Robolase IV; Chapter 7). 3. Individual motile sperm can be tracked in real-time (Robolase I; Chapters 2-3, 5-6). 4. While being tracked, velocity measurements are automatically calculated and saved (Robolase I; Chapters 2-3, 5-6). 5. Optical tweezers can be used to manipulate sperm and automatically calculate escape power using a power decay algorithm (Robolase I; Chapters 2-3, 5-6). 6. Labeled motile sperm can be analyzed in real-time with automated single or dual channel fluorescence imaging processing (Robolase I; Chapter 2-3).;Several experiments have been conducted using this platform to study sperm motility, fertilization, and early embryogenesis with applications to infertility and animal husbandry. This dissertation will describe the development of the automated optofluidic microscopy platform as well as the experiments performed.