Flow injection methods for drug-receptor interaction studies, based on probing cell metabolism

Abstract

The thesis describes the development of methods to study responses of living cells to chemical stimulation. To measure the responses, the developed methods utilize effects that the responses have on the rate of the cells' energy metabolism. The metabolic rate is probed either by measuring acid production or oxygen consumption. The technique of flow injection is used throughout the work to achieve swift and reproducible stimulant delivery and removal. Bead injection, a later variant of flow injection, is used to create optimal conditions for the detection of extracellular acidification and oxygen consumption. Detection of pH and [O2] is carried out by the appropriate fluorescent probes and a fluorescence microscope. The methods are validated by carbachol stimulation of Chinese Hamster Ovary M1 (CHO M1) cells.