Abstract

Nano scanning Auger microscopy (NanoSAM) and time-of-flight secondary ion mass spectrometry (TOF-SIMS) have been used in materials science research for some time, but NanoSAM, in particular, has only recently been applied to biological specimens. Here, the first concurrent utilization of NanoSAM, TOF-SIMS and microscopic techniques for the examination of a standard beverage fermentation strain of Saccharomyces pastorianus uncovered the presence of intracellular networks of CO2 in fermenting cells. Respiring cells produced few bubbles and instead had large internal vacuolar structures. Transmission electron microscopy analysis also showed osmiophilic layers at the cell exterior of fermenting cells that became more prevalent with fermentation duration, while osmiophilic layers were largely absent in respiring cells. TOF-SIMS analysis showed a compositional difference at the exterior and interior of SMA cells and between fermenting and respiring cells. Fermenting cells also appeared to have different 3-OH oxylipin profiles compared to respiring cells based upon examination with immunofluorescence microscopy. The results of this work and further study using these materials science techniques will substantially enhance our understanding of the chemical, ultrastructural and metabolic changes that occur in fermentation yeasts.

Highlights

  • A key tenet of materials science is that a comprehensive compositional survey of a substance is only possible when surface analysis is combined with depth profiling

  • At each of 24, 48 and 72 h, cells were collected from the 3 replicates of both growth mediums and were analyzed using light microscopy (LM), Nano scanning Auger microscopy (NanoSAM), high resolution (Hi-Res) scanning electron microscopy (SEM), transmission electron microscopy (TEM), TOF-SIMS and confocal laser scanning microscopy (CLSM)

  • Cells grown in yeast malt (YM) were often misshapen, whereas cells grown in yeast peptone glycerol (YPG) maintained a more ovoid shape

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Summary

Introduction

A key tenet of materials science is that a comprehensive compositional survey of a substance is only possible when surface analysis is combined with depth profiling. Comprehensive analysis of the SMA strain of Saccharomyces pastorianus. National Science and Engineering Research Council (NSERC) of Canada. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication

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