Fourier Transform Infrared Spectromicroscopy Research Articles

An infrared spectral biomarker accurately predicts neurodegenerative disease class in the absence of overt symptoms

Although some neurodegenerative diseases can be identified by behavioral characteristics relatively late in disease progression, we currently lack methods to predict who has developed disease before the onset of symptoms, when onset will occur, or the outcome of therapeutics. New biomarkers are needed. Here we describe spectral phenotyping, a new kind of biomarker that makes disease predictions based on chemical rather than biological endpoints in cells. Spectral phenotyping uses Fourier Transform Infrared (FTIR) spectromicroscopy to produce an absorbance signature as a rapid physiological indicator of disease state. FTIR spectromicroscopy has over the past been used in differential diagnoses of manifest disease. Here, we report that the unique FTIR chemical signature accurately predicts disease class in mouse with high probability in the absence of brain pathology. In human cells, the FTIR biomarker accurately predicts neurodegenerative disease class using fibroblasts as surrogate cells.

Addressable graphene encapsulation of wet specimens on a chip for optical, electron, infrared and X-ray based spectromicroscopy studies.

Label-free spectromicroscopy methods offer the capability to examine complex cellular phenomena. Electron and X-ray based spectromicroscopy methods, though powerful, have been hard to implement with hydrated objects due to the vacuum incompatibility of the samples and due to the parasitic signals from (or drastic attenuation by) the liquid matrix surrounding the biological object of interest. Similarly, for many techniques that operate at ambient pressure, such as Fourier transform infrared spectromicroscopy (FTIRM), the aqueous environment imposes severe limitations due to the strong absorption of liquid water in the infrared regime. Here we propose a microfabricated multi-compartmental and reusable hydrated sample platform suitable for use with several analytical techniques, which employs the conformal encapsulation of biological specimens by a few layers of atomically thin graphene. Such an electron, X-ray, and infrared transparent, molecularly impermeable and mechanically robust enclosure preserves the hydrated environment around the object for a sufficient time to allow in situ examination of hydrated bio-objects with techniques operating under both ambient and high vacuum conditions. An additional hydration source, provided by hydrogel pads lithographically patterned in the liquid state near/around the specimen and co-encapsulated, has been added to further extend the hydration lifetime. Note that the in-liquid lithographic electron beam-induced gelation procedure allows for addressable capture and immobilization of the biological cells from the solution. Scanning electron microscopy and optical fluorescence microscopy, as well as synchrotron radiation based FTIR and X-ray fluorescence microscopy, have been used to test the applicability of the platform and for its validation with yeast, A549 human carcinoma lung cells and micropatterned gels as biological object phantoms.

Compressed FTIR spectroscopy using low-rank matrix reconstruction.

Fourier transform infrared (FTIR) spectroscopy is a powerful technique in analytical chemistry. Typically, spatially distributed spectra of the substance of interest are conducted simultaneously using FTIR spectrometers equipped with array detectors. Scanning-based methods such as near-field FTIR spectroscopy, on the other hand, are a promising alternative providing higher spatial resolution. However, serial recording severely limits their application due to the long acquisition times involved and the resulting stability issues. We demonstrate that it is possible to significantly reduce the measurement time of scanning methods by applying the mathematical technique of low-rank matrix reconstruction. Data from a previous pilot study of Leishmania strains are analyzed by randomly selecting 5% of the interferometer samples. The results obtained for bioanalytical fingerprinting using the proposed approach are shown to be essentially the same as those obtained from the full set of data. This finding can significantly foster the practical applicability of high-resolution serial scanning techniques in analytical chemistry and is also expected to improve other applications of FTIR spectroscopy and spectromicroscopy.

Synchrotron FTIR spectromicroscopy as a tool for studying populations and individual living cells of green algae.

Fourier transform infrared (FTIR) spectromicroscopy was used to study individual living cells of three closely-related species of the green algal genus Chlamydomonas. Three types of spectral variation were observed between individual cells within a single culture, as well as between different cultures: variation around a mean, individual outliers, and the presence of subpopulations. By understanding and controlling this variation, we were able to spectroscopically differentiate between the three closely-related species. Spectral differences were confirmed using principal component analysis, leading to an understanding of the biochemical differences between species. This work highlights the additional information obtained by studying individual cells, and has implications for more traditional bulk measurements.

Supercontinuum-based Fourier transform infrared spectromicroscopy

Fourier-transform infrared (FTIR) spectromicroscopy combines the spatial resolution of optical microscopy with the spectral selectivity of vibrational spectroscopy. Synchrotron sources can provide diffraction-limited beams in the infrared, and therefore synchrotron-based FTIR spectromicroscopy is nowadays an indispensable tool for biology and materials science studies where high spatial resolution is required. However, the increasing need for accurate and highly spatially resolved characterization is calling for alternative laboratory-based sources to complement synchrotron radiation. To date, the low brightness of thermal emitters or high temporal coherence and narrow bandwidth or tunability of laser sources have hindered the progress of bench-top FTIR spectromicroscopy. Here, we demonstrate that fiber-based supercontinuum sources in the mid-infrared enable fast spectral mapping of localized material properties with close to diffraction-limited resolution (3 μm×3 μm) and pave the way to table-top, on-demand, fast, and highly spatially resolved studies. We illustrate these capabilities by imaging thin sections of human liver samples and compare the results and performance with those obtained using a synchrotron source.

The Temporal Degradation of Illicit Contaminants in Latent Fingermarks Using Fourier Transform Infrared Spectroscopic Imaging

Fourier Transform Infrared (FTIR) spectroscopy has been shown to be a rapid, non-destructive analytical technique capable of detecting trace amounts of exogenous particulate in fresh latent fingermarks. Fingermarks identified at a crime scene however, are rarely fresh and are likely to have been deposited days or even weeks before forensic analysis, leaving them prone to decomposition. In this study latent fingermarks were contaminated with trace amounts of the improvised explosive precursor’s ammonium nitrate and sodium chlorate, and the addictive narcotic cocaine. The latent fingermarks where then aged in natural daylight conditions over a 30 day period and intermittently analysed using FTIR spectromicroscopy. Exogenous particulate was identified using spectroscopic imaging of each fingermark, and spectra obtained from the samples were compared to control spectra to confirm the identification of the contaminant. This study demonstrates that these contaminants are detectable within latent fingermarks up to at least 30 days after deposition, with only ammonium nitrate showing signs of decomposition, and that fingermark chemistry has a minimal effect on their molecular integrity. This information has broad implications for the law enforcement community as it suggests that the detection of exogenous particulate within latent fingermarks is possible long after a suspect has handled an illegal substance. This study also confirms the use of spectroscopic imaging to provide a chemical signature for these illicit compounds weeks after deposition, and an image of the fingermark ridge pattern that can then be used for identification purposes.

Using new hetero-spectral two-dimensional correlation analyses and synchrotron-radiation-based spectromicroscopy to characterize binding of Cu to soil dissolved organic matter

Understanding the binding characteristics of copper (Cu) to different functional groups in soil dissolved organic matter (DOM) is important to explore Cu toxicity, bioavailability and ultimate fate in the environment. However, the methods used to explore such binding characteristics are still limited. Here, two-dimensional correlation spectroscopy (2DCOS) integrated with Fourier transform infrared (FTIR), 29Si nuclear magnetic resonance (NMR), 27Al NMR, and synchrotron-radiation-based FTIR spectromicroscopy were used to explore the binding characteristics of Cu to soil DOM as part of a long-term (23 years) fertilization experiment. Compared with no fertilization and inorganic fertilization (NPK), long-term pig manure fertilization (M) treatment significantly increased the concentration of total and bioavailable Cu in soils. Furthermore, hetero-spectral 2DCOS analyses demonstrated that the binding characteristics of Cu onto functional groups in soil DOM were modified by fertilization regimes. In the NPK treatment, Cu was bound to aliphatic C, whereas in the manure treatment SiO groups had higher affinity toward Cu than aliphatic C. Also, the sequence of binding of functional groups to Cu was modified by the fertilization treatments. Moreover, synchrotron-radiation-based FTIR spectromicroscopy showed that Cu, clay minerals and sesquioxides, and C functional groups were heterogeneously distributed at the micro-scale. Specifically, clay-OH as well as mineral elements had a distribution pattern similar to Cu, but certain (but not all) C forms showed a distribution pattern inconsistent with that of Cu. The combination of synchrotron radiation spectromicroscopy and 2DCOS is a useful tool in exploring the interactions among heavy metals, minerals and organic components in soils.

Opportunities for Live Cell FT-Infrared Imaging: Macromolecule Identification with 2D and 3D Localization

Infrared (IR) spectromicroscopy, or chemical imaging, is an evolving technique that is poised to make significant contributions in the fields of biology and medicine. Recent developments in sources, detectors, measurement techniques and speciman holders have now made diffraction-limited Fourier transform infrared (FTIR) imaging of cellular chemistry in living cells a reality. The availability of bright, broadband IR sources and large area, pixelated detectors facilitate live cell imaging, which requires rapid measurements using non-destructive probes. In this work, we review advances in the field of FTIR spectromicroscopy that have contributed to live-cell two and three-dimensional IR imaging, and discuss several key examples that highlight the utility of this technique for studying the structure and chemistry of living cells.

Multimode InfraRed Imaging and Microspectroscopy (MIRIAM) Beamline at Diamond

InfraRed (IR) MicroSpectroscopy is a quantitative analytical and non-destructive technique that has undergone a renaissance since its coupling to synchrotron radiation (SR) [1]. The primary advantages of an SR IR source with respect to BlackBody (BB) are a wide broadband spectrum, high brightness/brilliance, intrinsic light polarization, and temporal pulse structure. SR IR spans a larger spectral distribution, also covering the far-IR (or THz region), and can be up to 1000 times brighter in the mid-IR than thermal sources. Fourier Transform IR (FTIR) interferometry has proven extremely effective for performing IR absorption spectroscopy, i.e. for quantifying optically active vibrational modes of molecular components. SR's high brightness benefits FTIR spectromicroscopy with a signal-to-noise ratio unreachable by other broadband sources at diffraction limited spatial resolution at the microscopic scale and in confocal geometry.

Infrared spectromicroscopy of biochemistry in functional single cells

Over the years Fourier-Transform Infrared (FTIR) spectroscopy has been widely employed in the structural and functional characterization of biomolecules. The introduction of infrared (IR) microscopes and of synchrotron light sources has created expectations that FTIR could become a generally viable technique to study both structure and reactivity in vivo, inside single cells, by performing measurements that up to a few years ago were the preserve of in vitro experiments on purified macromolecules. In this review we present the state-of-the-art in the application of FTIR spectromicroscopy as a technique for the study of structure and dynamics in single cells, we discuss the performance requirements for this application and review developments in sample handling methods.

Identification of the finishing technique of an early eighteenth century musical instrument using FTIR spectromicroscopy

The study of varnishes from musical instruments presents the difficulty of analysing very thin layers of heterogeneous materials on samples most of which are generally brittle and difficult to prepare. Such study is crucial to the understanding of historical musical instrument varnishing practices since written sources before 1800 are very rare and not precise. Fourier-transform infrared (FTIR) spectroscopy and imaging methods were applied to identify the major chemical components within the build-up of the varnish layers on a cello made by one of the most prominent French violin-makers of the eighteenth century (Jacques Boquay, ca. 1680-1730). Two types of FTIR imaging methods were used: scanning with a synchrotron-based microscope and full-field imaging using a 2D imager with a conventional source. An interpretation of the results obtained from these studies on the Boquay cello is that the maker first applied a proteinaceous layer, probably gelatine-based animal glue. He later applied a second layer based on a mixture of a drying oil and diterpenic resin from Pinaceae sp. From an historical perspective, the results complement previous studies by describing a second technique used for musical instrument finishes at the beginning of the eighteenth century in Europe.

Fourier transform infrared evaluation of microscopic scarring in the cardiomyopathic heart: Effect of chronic AT 1 suppression

Our primary aim was to investigate the use of Fourier transform infrared (FTIR) spectromicroscopy as an accurate assay of cardiac extracellular matrix remodeling. Abnormal rearrangement or remodeling of the cardiac extracellular matrix is known to contribute to cardiac dysfunction. The microscopic multifocal necrosis and scarring are modulated by chronic AT 1 receptor blockade in experimental cardiomyopathy; thus, we also wished to rationalize the spectromicroscopic differences among control, untreated cardiomyopathic (CMP), and losartan-treated cardiomyopathic (LOS) hearts according to the pathogenesis of experimental cardiomyopathy. Male UM-X7.1 cardiomyopathic Syrian hamsters at early and late (65 and 200 days) stages of cardiomyopathy were subjected to 4-week losartan (15 mg/kg/day continuous infusion) treatment. Focal collagen microdomain distribution was confirmed spectroscopically by observation of the collagen IR fingerprint in the 1000–1800 cm −1 region. Synchrotron FTIR spectromicroscopic map data were obtained from control (F1-β strain) hamsters, nontreated cardiomyopathic, and losartan-treated CMP animals and imaged with mapping software, according to intensity of collagen fingerprint. Compared to controls, untreated late-stage CMP myocardium was characterized by elevated levels of fibrillar collagens and this was partially normalized with a 4-week losartan treatment. FTIR spectromicroscopy revealed that elevated collagen expression in focal microdomains is present in late-stage cardiomyopathy, and 4-week AT 1 blockade is associated with attenuation of collagen absorption in these lesions.