Probing Bacterial Stress Responses from Energetic Perspective(3/4)

The aim of this project is to probe bacterial stress responses from energetic respective. There are three objectives. 1. Developing single cell high-throughput and high resolution optical measurement systems. 2. Developing cutting edge single cell energetic measurements and optoenergetic tools. 3. To study bacterial energetic survival kinetics..Cellular energetics is a developing biophysics fields combining single cell precise measurement and high throughput data collection to expand us understand to the single cell survival dynamics. We can investigate single cell functional mechanism and cell-to-cell variation. It is a new field standing on the giant’s shoulder that we are going to developing a serious technique to study bacterial energetic responses to the external stresses.A cell is a fundamental unit of life. Single cell bacterium is about few micro-meter in size and facing hostile environments. Bacterial cells in natural environments are typically in the “feast or famine” status seeking their survival. They can grow rapidly encountering resources and maintain necessities under stresses. We have good understanding of cell growth biochemistry, physiology and cell division leading to exponential growth. However, the mechanisms of cellular energetic stress response and the exponential decay death kinetics are limited. Material, information and energy are the three fundamental elements of living organisms. We have fruitful knowledge of biological materials and the information storage/transduction. However, there is no comprehensive understanding of cellular energetics. In order to reveal the usage and distribution of bacterial cellular energetics, we propose the following research. Firstly, we are going to develop single cell high-throughput and high-resolution optical measurement system. Traditional high-throughput experiments are limited for low resolution measurements. For example, flow cytometry can only provide low spatial resolution information from cells. On the other hands, high resolution experiments such as super-resolution fluorescence microscopy can only target limit number of cells at the same time. Therefore, there is an urgent need to develop microfluidic devices achieving both high-throughput and high-resolution experiments. Secondly, we will continue to develop single-cell physiology/energetic measuring tools. We have developed single-cell intracellular pH and using bacterial flagellar motor rotation to measure the PMF. To probe the bacterial cellular energetic response, we are going to develop new membrane potential probes, improving the ATP sensing protein Queen in bacteria and optical proton pumps. Thirdly, our final goal is to understand bacterial energetic consumption and survival kinetics. Combining the new experimental systems and energetic parameters measurements, we are able to study the bacterial energy status, consumption and survival kinetics. Though gained new knowledge, we will be able to have new perspective to understand bacterial survival and potentially, a new solution for the anti-superbug war.