Recent measurement methods for population and single cell dynamics

The purpose of this work is to develop laboratory techniques that allow quantitative measurements of bacterial growth and expression. An introductory section was written to prepare the reader for the contents of this work. In the first part, bulk measurements of batch cultures are evaluated. A low cost machine designed for this purpose is presented. We found that the measurements taken by this machine are comparable to those taken more expensive equipment. Later on, more elaborated real time measurements of bacterial growth curves are documented for 5 different strain of Escherichia coli MG1655. Changes in growth rate were linked to the expression of specific reporter proteins in each strain. In the second part, single cell measurements are assessed. Microfluidic assisted cell sorting (MACS) and flow cytometry (FC) are compared as techniques to study bacterial morphology and gene expression. MACS was found to yield higher quality data, in spite of not having as high throughput as FC. Advances towards mother machine experiments were done. Image and data analysis software was designed as well as software for controlling hardware available in the laboratory. Induction of bacterial biofilms was achieved inside of mother machine chips. The collection of methods described in this work can serve as a guide for experimental systems biology for new members in the biophysics laboratory at Universidad de los Andes

• Autores: Sánchez Isaza, Carlos Arturo
• Tipo de recurso: Trabajo de grado de pregrado
• Fecha de publicación: 2017
• Institución: Universidad de los Andes
• Repositorio: Séneca: repositorio Uniandes
• Idioma: eng
• OAI Identifier: oai:repositorio.uniandes.edu.co:1992/39778
• Acceso en línea: http://hdl.handle.net/1992/39778
• Palabra clave: Biofísica; Microfluídica; Expresión génica; Escherichia coli; Física
• Rights: openAccess
• License: http://creativecommons.org/licenses/by-nc-sa/4.0/