Alessandra Masini

X-ray irradiation of yeast cells

Saccharomyces Cerevisiae yeast cells were irradiated using the soft X-ray laser-plasma source at Rutherford Laboratory. The aim was to produce a selective damage of enzyme metabolic activity at the wall and membrane level (responsible for fermentation) without interfering with respiration (taking place in mitochondria) and with nuclear and DNA activity. The source was calibrated by PIN diodes and X-ray spectrometers. Teflon stripes were chosen as targets for the UV laser, emitting X-rays at about 0.9 keV, characterized by a very large decay exponent in biological matter. X-ray doses to the different cell compartments were calculated following a Lambert-Bouguet-Beer law. After irradiation, the selective damage to metabolic activity at the membrane level was measured by monitoring CO2 production with pressure silicon detectors. Preliminary results gave evidence of pressure reduction for irradiated samples and non-linear response to doses. Also metabolic oscillations were evidenced in cell suspensions and it was shown that X-ray irradiation changed the oscillation frequency.

Biosystem response to soft-X-rays irradiation: non-monotonic effects in the relevant biological parameters of yeast cells

SummarySoft-X-rays irradiation of yeast cells allows selective interference with different cellular structures. The monitoring of different physical parameters leads to substantial variations in the response to X-rays showing that monotonicity should not be taken for granted.

An ensemble of new techniques to study soft-X-ray-induced variations in cellular metabolism

An ensemble of new techniques has been developed to study cell metabolism. These include: CO2 production monitoring, cell irradiation with soft X rays produced with a laser-plasma source, and study of oscillations in cell metabolic activity via spectral analysis of experimental records. Soft X-rays at about 0.9 keV, with a very low penetration in biological material, were chosen to produce damages at the metabolic level, without great interference with DNA activity. The use of a laser-plasma source allowed a fast deposition of high doses. Monitoring of CO2 production allowed us to measure cell metabolic response immediately after irradiation in a continuous and non invasive way. Also a simple model was developed to calculate X-ray doses delivered to the different cell compartments following a Lambert-Bouguet–Beer law. Results obtained on Saccharomyces cerevisiae yeast cells in experiments performed at Rutherford Appleton Laboratory are presented.

Contact x-ray microscopy using Asterix

The use of a high energy laser source for soft x-ray contact microscopy is discussed. Several different targets were used and their emission spectra compared. The x-ray emission, inside and outside the Water Window, was characterized in detail by means of many diagnostics, including pin hole and streak cameras. Up to 12 samples holders per shot were exposed thanks to the large x-ray flux and the geometry of the interaction chamber. Images of several biological samples were obtained, including Chlamydomonas and Crethidia green algae, fish and boar sperms and Saccharomyces Cerevisiae yeast cells. A 50 nm resolution was reached on the images of boar sperm. Original information concerning the density of inner structures of Crethidia green algae were obtained.

Soft x-ray-controlled dose deposition in yeast cells: techniques, model, and biological assessment

A procedure is presented to release soft x-rays onto yeast cell membrane allegedly damaging the resident enzymatic processes connected with fermentation. The damage is expected to be restricted to regulating fermentation processes without interference with respiration. By this technique fermentation is followed leading to CO2 production, and respiration resulting in global pressure measurements. A solid state pressure sensor system has been developed linked to a data acquisition system. Yeast cells cultures have been investigated at different concentrations and with different nutrients. A non-monotone response in CO2 production as a function of the delivered x-ray dose is observed.

Measurement and Characterization of Cellular Metabolic Activity by CO2 Production Monitoring, Soft X-ray Irradiation and Advanced Spectral Analysis

Results obtained using a new technique for studying cell metabolism are presented. The technique, consisting in CO2 production monitoring, has been applied to Saccharomyces cerevisiae yeast cells. Also, the cells were irradiated using soft X-ray generated by a laser-plasma source with the aim of producing a damage of metabolic processes at the wall level. X-ray doses delivered to the different cell compartments were estimated using a Lambert-Bouguet-Beer law. Immediately after irradiation, the effects on metabolic activity were measured by monitoring CO2 production. This has evidenced oscillations in cell metabolism and X-ray induced changes in oscillation frequency. In order to capture the main frequency components of the oscillation, minimum bias techniques has been used for spectral estimates.