The Potential of Confocal Imaging for Measuring Physiological Changes in Brewer's Yeast

Schlee, Cornelia; Miedl, Michaela; Leiper, Kenneth A.; Stewart, Graham G.

doi:10.1002/j.2050-0416.2006.tb00243.x

Cited by 14 publications

(8 citation statements)

References 75 publications

(66 reference statements)

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“…Functional and structural investigations of the plasma membrane are necessary to understand the mechanisms operating during dehydration perturbation. To this end, we used fluorescence microscopy (epifluorescence and confocal) to observe modifications of the yeast plasma membrane during dehydration [25]. These modifications included changes in plasma membrane permeability, Sur7-GFP (contained in MCC) repartition, and membrane delocalization.…”

Section: Introductionmentioning

confidence: 99%

Lateral reorganization of plasma membrane is involved in the yeast resistance to severe dehydration

Dupont¹,

Beney²,

Ritt³

et al. 2010

Biochimica et Biophysica Acta (BBA) - Biomembranes

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In this study, we investigated the kinetic and the magnitude of dehydrations on yeast plasma membrane (PM) modifications because this parameter is crucial to cell survival. Functional (permeability) and structural (morphology, ultrastructure, and distribution of the protein Sur7-GFP contained in sterol-rich membrane microdomains) PM modifications were investigated by confocal and electron microscopy after progressive (non-lethal) and rapid (lethal) hyperosmotic perturbations. Rapid cell dehydration induced the formation of many PM invaginations followed by membrane internalization of low sterol content PM regions with time. Permeabilization of the plasma membrane occurred during the rehydration stage because of inadequacies in the membrane surface and led to cell death. Progressive dehydration conducted to the formation of some big PM pleats without membrane internalization. It also led to the modification of the distribution of the Sur7-GFP microdomains, suggesting that a lateral rearrangement of membrane components occurred. This event is a function of time and is involved in the particular deformations of the PM during a progressive perturbation. The maintenance of the repartition of the microdomains during rapid perturbations consolidates this assumption. These findings highlight that the perturbation kinetic influences the evolution of the PM organization and indicate the crucial role of PM lateral reorganization in cell survival to hydric perturbations.

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Section: Introductionmentioning

confidence: 99%

Lateral reorganization of plasma membrane is involved in the yeast resistance to severe dehydration

Dupont¹,

Beney²,

Ritt³

et al. 2010

Biochimica et Biophysica Acta (BBA) - Biomembranes

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“…In particular, the expression and sub cellular localisation of proteinase A during yeast fermentation has been studied employing a green fluorescent protein clone 117 . These studies confirm previous findings 13 , using the Kondo et al assay 67 method for proteinase A, which demonstrated that this enzyme is located primarily in the yeast vacuole, but under stress conditions significant activity appears in the cell cytoplasm.…”

Section: Confocal Microscopymentioning

confidence: 99%

“…A number of physiological parameters and cell compounds in yeast cells (glycogen, neutral lipids, trehalose, bud scars, DNA and intracellular proteinases) have been successfully visualised with the aid of highly specific fluorochromes. In particular, the expression and sub cellular localisation of proteinase A during yeast fermentation has been studied employing a green fluorescent protein clone 117 . These studies confirm previous findings 13 , using the Kondo et al assay 67 method for proteinase A, which demonstrated that this enzyme is located primarily in the yeast vacuole, but under stress conditions significant activity appears in the cell cytoplasm.…”

Section: Confocal Microscopymentioning

confidence: 99%

The Horace Brown Medal Lecture: Forty Years of Brewing Research

Stewart

2009

Journal of the Institute of Brewing

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Horace Brown spent fifty years conducting brewing research in Burton-on-Trent, Dublin and London. His contributions were remarkable and his focus was to solve practical brewing problems by employing and developing fundamental scientific principles. He studied all aspects of the brewing process including raw materials, wort preparation, fermentation, yeast and beer stability. As a number of previous presenters of the Horace Brown Lecture have discussed Brown's achievements in detail, the focus of this paper is a review of the brewing research that has been conducted by the author and his colleagues during the past forty years. Similar to Horace Brown, fundamental research has been employed to solve brewing problems. Research studies that are discussed in this review paper include reasons for premature flocculation of ale strains resulting in wort underattenuation including mechanisms of co-flocculation and pure strain flocculation, storage procedures for yeast cultures prior to propagation, studies on the genetic manipulation of brewer's yeast strains with an emphasis on the FLO1 gene, spheroplast fusion and the respiratory deficient (petite) mutation, the uptake and metabolism of wort sugars and amino acids, the influence of wort density on fermentation characteristics and beer flavour and stability, and finally, the contribution that high gravity brewing has on brewing capacity, fermentation efficiency and beer quality and stability.

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“…There was done a lot of work on Saccharomyces cerevisiae yeast using fluorescence microscopy. Valuable review of LSCM applied on yeasts S. cerevisiae is summarized in (7). This review is dealing with staining both glycogen and neutral lipids as a storage material, staining inner membranes (like vacuolar membrane), nucleic acids and bud scars.…”

Section: Introductionmentioning

confidence: 99%

Fluorescence lifetime imaging of red yeast Cystofilobasidium capitatum during growth

Vanek¹,

Mravec²,

Szotkowski³

et al. 2018

The EuroBiotech Journal

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Red yeast Cystofilobasidium capitatum autofluorescence was studied by means of confocal laser scanning microscopy (CLSM) to reveal distribution of carotenoids inside the cells. Yeasts were cultivated in 2L fermentor on glucose medium at permanent light exposure and aeration. Samples were collected at different times for CLSM, gravimetric determination of biomass and HPLC determination of pigments. To compare FLIM (Fluorescence Lifetime Imaging Microscopy) images and coupled data (obtained by CLSM) with model systems, FLIM analysis was performed on micelles of SDS:ergosterol and SDS:coenzyme Q with different content of ergosterol and coenzyme Q, respectively, and with constant addition of beta-carotene. Liposomes lecithin:ergosterol:beta-carotene were investigated too. Two different intracellular forms of carotenoids were observed during most of cultivations, with third form appeared at the beginning of stationary phase. Observed behavior is probably due to formation of some kind of carotenoid protective system in membranes of different compartments of yeast cell, especially cytoplasmic membrane.

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The Potential of Confocal Imaging for Measuring Physiological Changes in Brewer's Yeast

Cited by 14 publications

References 75 publications

Lateral reorganization of plasma membrane is involved in the yeast resistance to severe dehydration

Lateral reorganization of plasma membrane is involved in the yeast resistance to severe dehydration

The Horace Brown Medal Lecture: Forty Years of Brewing Research

Fluorescence lifetime imaging of red yeast Cystofilobasidium capitatum during growth

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