FT-IR microspectroscopy for microbiological studies

Orsini, F.; Ami, Diletta; Villa, Anna; Sala, G; Bellotti, M.; Doglia, S.

doi:10.1016/s0167-7012(00)00168-8

Cited by 93 publications

(67 citation statements)

References 14 publications

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“…Absorption spectra with a low noise level were obtained by setting the microscope aperture at about 100 lm · 100 lm [9]. The background spectrum was collected before each measurement and no baseline correction was required on the spectra.…”

Section: Ft-ir Microspectroscopymentioning

confidence: 99%

Kinetics of inclusion body formation studied in intact cells by FT‐IR spectroscopy

et al. 2005

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The aggregation of a recombinant lipase as inclusion bodies (IBs) was studied directly within intact Escherichia coli cells by FT-IR microspectroscopy. Through this approach, it was possible to monitor in real time the different kinetics of IB formation at 37 and 27°C, in excellent agreement with the results of the SDS-PAGE analysis. Furthermore, insights on the residual native-like structure of the expressed protein within IB -both isolated and inside cells -were obtained by the secondary structure analysis of the Amide I band in the IB FT-IR spectra.

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Section: Ft-ir Microspectroscopymentioning

confidence: 99%

Kinetics of inclusion body formation studied in intact cells by FT‐IR spectroscopy

et al. 2005

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“…Two main different types of infrared microscopy can be used: the first, conventional, allows the collection of the IR absorption spectra from a microvolume within the sample, with a spatial resolution not only due to the diffraction limit of Mid IR light (3-10 μm), but also to the level of the absorbed light. For these reasons, a spectrum of good quality can be collected by an area larger than 20 μm x 20 μm -when using a nitrogen cooled Mercury Cadmium Telluride (MCT) detector (Orsini et al, 2000). Thanks to the variable aperture of the microscope, it is possible to select a small area of few tens of microns within the sample, enabling the study of intact cells (Tanthanuc et al, 2010;Thumanu et al, 2009;Ami et al, 2008;Wood et al, 2008), tissues (Choo et al, 1996) and whole model organisms, as nematodes (Diomede et al, 2010;Ami et al, 2004).…”

Section: Infrared Microspectroscopy Applied To the Study Of Intact Cementioning

confidence: 99%

“…Typically, measurements on biomolecules and on complex biological systems, such as cells and tissues, are carried out in transmission, employing suitable IR transparent supports, for instance of barium fluoride or zinc selenide. However, sometimes, it could be useful to work in ATR when the samples of interest are highly absorbing or when they cannot be easily transferred onto an opportune infrared support (Walsh et al, 2007;Orsini et al, 2000). Indeed, in ATR measurements the sample is placed in contact with the ATR element (diamond, germanium, etc) characterized by a refractive index higher than that of the sample.…”

Section: Infrared Microspectroscopy Applied To the Study Of Intact Cementioning

confidence: 99%

Fourier Transform Infrared Microspectroscopy as a Tool for Embryonic Stem Cell Studies

Ami¹,

Mereghetti²,

Natalello³

et al. 2011

Methodological Advances in the Culture, Manipulation and Utilization of Embryonic Stem Cells for Basic and Practical Applicatio

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“…This method has been extensively used in recent years for the study of microbial surfaces, and the identification and classification of microorganisms [20][21][22][23].…”

Section: Introductionmentioning

confidence: 99%

Li+ effect on the cell wall of the yeast Saccharomyces cerevisiae as probed by FT-IR spectroscopy

2013

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+ effect on the cell wall of the yeast Saccharomyces cerevisiae as probed by FT-IR spectroscopy IntroductionThe capacity of metal cations to induce competence of yeast and bacterial cells to exogenous DNA is the basis of biological methods in biotechnology and molecular genetics [1][2][3]. The most commonly used, efficient protocol, for genetic transformation of Saccharomyces cerevisiae is treatment with alkali metal ions. Li + ions are the most effective of all the cations tested, and the transformation efficiency is comparable to levels obtained by the protoplast method [4]. Li + ions enhance the transformation of intact cells, but no effect is observed on transformation of protoplasts, implying that Li + ions facilitate DNA passage through the cell wall [5]. The mechanism underlying S. cerevisiae transformation includes DNA attachment and penetration through the cell wall, although how DNA passes through the cell wall is not yet clear [6][7][8]. Cell wall density, thickness and structure are factors of major importance during penetration of exogenous molecules into the cell. However, despite the extensive use of Li + ions in yeast transformation protocols, the influence of these cations on the structure of the yeast cell wall has not been widely investigated and, therefore, remains unclear. The influence of Li + ions on the cell surface topography of intact S. cerevisiae cells was observed by atomic force microscopy and it was found that the surface of Li + treated yeast cells became much rougher [9].The S. cerevisiae cell wall is composed of three major components: β-glucans, chitin, and mannoproteins. Glucose residues are linked to other glucose molecules through β(1→3) and β(1→6) linkages and to N-acetylglucosamine via β(1→4) bonds [10,11]. S. cerevisiae mannan has a linear α(1→6)-linked Keywords: FT-IR spectroscopy • Lithium • Saccharomyces cerevisiae • TransformationAbstract: The effect of Li + ions as a transformation inducing agent on the yeast cell wall has been studied. Two Saccharomyces cerevisiae strains, p63-DC5 with a native cell wall, and strain XCY42-30D(mnn1) which contains structural changes in the mannan-protein complex, were used. Fourier transform infrared (FT-IR) spectroscopy has been used for the characterization of the yeast strains and for determination of the effect of lithium cations on the cell wall. A comparison of the carbohydrate absorption band positions in the 970-1185 cm -1 range, of Na + and Li + treated yeast cells has been estimated. Absorption band positions of the cell wall carbohydrates of p63-DC5 were not influenced by the studied ions. On the contrary, the treatment of XCY42-30D(mnn1) cells with Li + ions shifted glucan band positions, implying that the cell wall structure of strain XCY42-30D(mnn1) is more sensitive to Li + ion treatment. backbone with side chains of α(1→2)-and α(1→3)-linked mannose units. In S. cerevisiae, mannoproteins contribute to the regulation of cell wall porosity, and therefore control both the exit of secretary proteins and the entrance of ma...

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FT-IR microspectroscopy for microbiological studies

Cited by 93 publications

References 14 publications

Kinetics of inclusion body formation studied in intact cells by FT‐IR spectroscopy

Kinetics of inclusion body formation studied in intact cells by FT‐IR spectroscopy

Fourier Transform Infrared Microspectroscopy as a Tool for Embryonic Stem Cell Studies

Li+ effect on the cell wall of the yeast Saccharomyces cerevisiae as probed by FT-IR spectroscopy

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