ИСТИНА

Bioluminescence is convenient and sensitive instrument for monitoring of different biological processes in living organisms. The advantage of a firefly luciferase – luciferin system is that it allows to detect several parameters at once – ATP (or metabolic activity), luciferase activity (luminescent marker), Mg2+ ions and influence of O2. Intracellular ATP is universal indicator for the express monitoring of metabolic activity of living cells. It is a rapid marker for studying of mechanisms of cell stress and viability. The system with destruction of cells by DMSO with the subsequent detection of ATP in the received extract is used. DMSO has the following advantages over lytic agents of other classes – first, it stops immediately all intracellular processes, including synthesis and hydrolysis of ATP and, thus, the true content of ATP measurement in cell is achieved. Secondly, spontaneous hydrolysis of ATP in DMSO practically absent and extracts can be stored for a long time before analysis. In the third, high stability of a bioluminescent signal at ATP measurement is observed. Another indicator is the ratio of intracellular and extracellular ATP. It characterizes a cell lysis degree – increase in content of extracellular ATP is a result of a permeabilization or destruction of a cell membrane. The method is useful for studying of cell death and stress effects of the physical and chemical nature, cytotoxicity, hemolysis, for studying of mechanisms of resistance of cells to bioactive agents, for screening of medicines ets.