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The air sensitivity of many substrates, and specifically biosurfaces, presents an experimental challenge for their analysis by vibrational spectroscopy and, in particular, infrared microscopy on a nanometer scale. The recent development of atomic-force-microscopy-based infrared spectroscopy (AFM-IR), which circumvents the Abbe diffraction limit, allows nanoscale chemical characterization of surfaces. Additionally, this technique has been shown to work for thin films under aqueous environments but is limited to substrates up to 10 nm thick, thus ruling out application to many biological surfaces. To circumvent this restriction, we have utilized hydrogels to cover such surfaces and maintain a more physiologically representative environment for biological substrates. We show that it is feasible to use AFM-IR to chemically characterize this type of substrate buried under a thin hydrogel film. Specifically, this work describes the AFM-IR spectra of red blood cells under polyvinyl alcohol hydrogels., (Copyright © 2020 Biophysical Society. Published by Elsevier Inc. All rights reserved.)

The challenge of deriving quantitative information from the infrared spectra of proteins arises from the large number of secondary structures and amino acid side-chain functional groups that all contribute to the spectral intensity, such as within the amide I band (1600-1700 cm -1 ). The band is invariably heavily convoluted from overlapping spectral features, thereby making interpretation difficult such that deconvolution is usually required. This work critically examines the methods available to deconvolute the spectra and assesses the commonly used methods and algorithms applied to vibrational spectra for smoothing and peak identification. We show that unless their spectra have very high signal-to-noise ratios, quantitative analysis to decipher protein constituents is not feasible. The advantages and disadvantages of spectral smoothing using adjacent averaging, the Savitzky-Golay filter and the fast Fourier transform filter are examined in detail. The use of derivative spectra to identify peaks is described with particular reference to the influence and reduction of interfering water bands in the amide I region. The reliability of band narrowing techniques such as second-derivative analysis or Fourier deconvolution that lead to the identification of the contributing protein peaks is investigated. Both methods are shown to be limited in their capacity to resolve features with very similar frequencies. Additionally, the presence of narrow bands arising from high-frequency noise whether from atmospheric water vapor, acoustic vibrations, or electrical interference results in both methods becoming increasingly unusable as narrow bands are preferentially enhanced at the expense of broad ones such as the amide I bands. An optimal strategy is critically developed to allow accurate determination and quantification of protein constituents and their conformations. Additionally, quantitative methods are proposed to account for baseline shifts, which would otherwise introduce significant errors in similarity indices.

Molecular scale information about the structure of surfactants at interfaces underlies their application in consumer products. In this study the non-linear optical technique of Sum Frequency Generation (SFG) vibrational spectroscopy has been used to investigate the structure and temporal behaviour of two cationic surfactants used frequently in hair conditioners. SFG spectra of films of behenyltrimethylammonium methosulfate (BTMS) and behenyltrimethylammonium chloride (BTAC) were recorded at the air/water interface and on glass slides following Langmuir Blodgett (LB) deposition. The assignment of the BTMS and BTAC spectral features (resonances) to the CH stretching modes of the surfactants was consolidated by comparison with the SFG spectrum of deuterated cetyltrimethylammonium bromide (d-CTAB) and by recording spectra on D 2 O as well as on water. The CH resonances arise from the methylene and methyl groups of the tail and head-groups of the surfactants. A slow collapse mechanism was observed following film compression of both BTAC and BTMS. The change in molecular structure of the films undergoing this slow collapse was followed by recording sequential SFG spectra in the CH region, and by monitoring the SFG intensity at specific wavenumbers over time. Additionally, LB deposition onto glass was used to capture the state of the film during the slow collapse, and these SFG spectra showed close similarity to the corresponding spectra on water. Complementary Atomic Force Microscopy (AFM) was used to elucidate the layering of the compressed and relaxed films deposited onto mica by LB deposition., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Superoxide-based nonaqueous metal-oxygen batteries have received considerable research attention as they exhibit high energy densities and round-trip efficiencies. The cycling performance, however, is still poor. Here we study the cycling characteristic of a Na-O 2 battery using solid-state nuclear magnetic resonance, Raman spectroscopy, and scanning electron microscopy. We find that the poor cycling performance is primarily caused by the considerable side reactions stemming from the chemical aggressiveness of NaO 2 as both a solid-phase and dissolved species in the electrolyte. The side reaction products cover electrode surfaces and hinder electron transfer across the electrode-electrolyte interface, being a major reason for cell failure. In addition, the available electrode surface and porosity change considerably during cell discharging and charging, affecting the diffusion of soluble species (superoxide and water) and resulting in inhomogeneous reactions across the electrode. This study provides insights into the challenges associated with achieving long-lived superoxide-based metal-O 2 batteries.