Casting new physicochemical light on the fundamental biological processes in single living cells by using Raman microspectroscopy.

This Personal Account highlights the capabilities of spontaneous Raman microspectroscopy for studying fundamental biological processes in a single living cell. Raman microspectroscopy provides time- and space-resolved vibrational Raman spectra that contain detailed information on the structure and dynamics of biomolecules in a cell. By using yeast as a model system, we have made great progress in the development of this methodology. The results that we have obtained so far are described herein with an emphasis placed on how three cellular processes, that is, cell-division, respiration, and cell-death, are traced and elucidated with the use of time- and space-resolved structural information that is extracted from the Raman spectra. For cell-division, compositional- and structural changes of various biomolecules are observed during the course of the whole cell cycle. For respiration, the redox state of mitochondrial cytochromes, which is inferred from the resonance Raman bands of cytochromes, is used to evaluate the respiration activity of a single cell, as well as that of isolated mitochondrial particles. Special reference is made to the "Raman spectroscopic signature of life", which is a characteristic Raman band at 1602 cm(-1) that is found in yeast cells. This signature reflects the cellular metabolic activity and may serve as a measure of mitochondrial dysfunction. For cell-death, "cross-talk" between the mitochondria and the vacuole in a dying cell is suggested.