Nanoscience and Nanotechnology CONFOCAL MICROSCOPY AND ATOMIC FORCE MICROSCOPY OF SACCHAROMYCES CEREVISIAE (original) (raw)

We examined the physical properties of the surrounding yeast cell walls by using atomic force microscopy (AFM). The yeast cells were prepared on a cleaned glass substrate for CM observation and were mechanically trapped into a porous membrane for AFM measurement. The confocal image of the yeast cells was measured in air meanwhile the AFM topography images of the cells were measured in both de-ionized (DI) water (pH = 6.9) and phosphate-buffered saline (PBS) solution (pH = 7.4). No significant differences between the AFM topography images of the yeast cells measured in DI water and in phosphate buffered saline (PBS) solution could be inferred. In order to get the quantitative information on the sample elasticity, the force curves between an AFM tip and the yeast cell have been measured. These curves were measured in both DI water and in PBS solution on the same yeast cell using the same AFM cantilever to get the reliable result. The contact region of these force curves in approach mode was then converted into force versus indentation curves. These curves would be fitted with Hertz-Sneddon model for the calculation of the elasticity. Analysis of the curves indicates that there is a difference of the Young's modulus values of the yeast cell in various environments. These data show that the salt buffer solution increases the rigidity of the biological system.

An Atomic Force Microscopy study of yeast response to ethanol stress: Evidence for a role of the plasma membrane in the nanomechanical properties of the cell walls

Applied and Environmental Microbiology, 2016

ABSTRACTA wealth of biochemical and molecular data have been reported regarding ethanol toxicity in the yeastSaccharomyces cerevisiae. However, direct physical data on the effects of ethanol stress on yeast cells are almost nonexistent. This lack of information can now be addressed by using atomic force microscopy (AFM) technology. In this report, we show that the stiffness of glucose-grown yeast cells challenged with 9% (vol/vol) ethanol for 5 h was dramatically reduced, as shown by a 5-fold drop of Young's modulus. Quite unexpectedly, a mutant deficient in the Msn2/Msn4 transcription factor, which is known to mediate the ethanol stress response, exhibited a low level of stiffness similar to that of ethanol-treated wild-type cells. Reciprocally, the stiffness of yeast cells overexpressingMSN2was about 35% higher than that of the wild type but was nevertheless reduced 3- to 4-fold upon exposure to ethanol. Based on these and other data presented herein, we postulated that the ef...

Evidence for a Role for the Plasma Membrane in the Nanomechanical Properties of the Cell Wall as Revealed by an Atomic Force Microscopy Study of the Response of Saccharomyces cerevisiae to Ethanol Stress

Applied and Environmental Microbiology, 2016

A wealth of biochemical and molecular data have been reported regarding ethanol toxicity in the yeast Saccharomyces cerevisiae . However, direct physical data on the effects of ethanol stress on yeast cells are almost nonexistent. This lack of information can now be addressed by using atomic force microscopy (AFM) technology. In this report, we show that the stiffness of glucose-grown yeast cells challenged with 9% (vol/vol) ethanol for 5 h was dramatically reduced, as shown by a 5-fold drop of Young's modulus. Quite unexpectedly, a mutant deficient in the Msn2/Msn4 transcription factor, which is known to mediate the ethanol stress response, exhibited a low level of stiffness similar to that of ethanol-treated wild-type cells. Reciprocally, the stiffness of yeast cells overexpressing MSN2 was about 35% higher than that of the wild type but was nevertheless reduced 3- to 4-fold upon exposure to ethanol. Based on these and other data presented herein, we postulated that the effect...

Analytical Models for Measuring the Mechanical Properties of Yeast

Cells

The mechanical properties of yeast play an important role in many biological processes, such as cell division and growth, maintenance of internal pressure, and biofilm formation. In addition, the mechanical properties of cells can indicate the degree of damage caused by antifungal drugs, as the mechanical parameters of healthy and damaged cells are different. Over the past decades, atomic force microscopy (AFM) and micromanipulation have become the most widely used methods for evaluating the mechanical characteristics of microorganisms. In this case, the reliability of such an estimate depends on the choice of mathematical model. This review presents various analytical models developed in recent years for studying the mechanical properties of both cells and their individual structures. The main provisions of the applied approaches are described along with their limitations and advantages. Attention is paid to the innovative method of low-invasive nanomechanical mapping with scanning...

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