Towards practical soft X-ray spectromicroscopy of biomaterials

Hitchcock, Adam P.; Morin, Cynthia; Heng, Y. M.; Cornelius, Rena M.; Brash, John L.

doi:10.1163/156856202320401960

Cited by 73 publications

(65 citation statements)

References 26 publications

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“…Figure 4 presents the C 1s spectra of albumin (protein), alginate (polysaccharide), 1-palmitoyl-2-hydroxy-sn-glycero-3-phosphocholine (saturated lipid), and calf thymus DNA. The spectrum of albumin (12,26) is similar to that of fibrinogen (12) and other proteins (43), as expected from the averaging over the C 1s spectra of the individual amino acids (16). The spectrum of DNA is qualitatively in agreement with that reported by Zhang et al (43), but the energy scale differs by 0.3 eV.…”

Section: Resultssupporting

confidence: 82%

Scanning Transmission X-Ray, Laser Scanning, and Transmission Electron Microscopy Mapping of the Exopolymeric Matrix of Microbial Biofilms

Lawrence¹,

Swerhone²,

Leppard³

et al. 2003

Appl Environ Microbiol

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327

270

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Confocal laser scanning microscopy (CLSM), transmission electron microscopy (TEM), and soft X-ray scanning transmission X-ray microscopy (STXM) were used to map the distribution of macromolecular subcomponents (e.g., polysaccharides, proteins, lipids, and nucleic acids) of biofilm cells and matrix. The biofilms were developed from river water supplemented with methanol, and although they comprised a complex microbial community, the biofilms were dominated by heterotrophic bacteria. TEM provided the highestresolution structural imaging, CLSM provided detailed compositional information when used in conjunction with molecular probes, and STXM provided compositional mapping of macromolecule distributions without the addition of probes. By examining exactly the same region of a sample with combinations of these techniques (STXM with CLSM and STXM with TEM), we demonstrate that this combination of multimicroscopy analysis can be used to create a detailed correlative map of biofilm structure and composition. We are using these correlative techniques to improve our understanding of the biochemical basis for biofilm organization and to assist studies intended to investigate and optimize biofilms for environmental remediation applications.Scanning transmission X-ray microscopy (STXM) (1, 2), which uses near-edge X-ray absorption spectroscopy (NEXAFS) as its contrast mechanism, is a powerful new tool that can be applied to fully hydrated biological materials. This is possible due to the ability of soft X rays to penetrate water, the presence of suitable analytical core edges in the soft X-ray region, and reduced radiation damage (compared to that caused by electron beam techniques). Soft X rays also provide spatial resolution of better than 50 nm, which is suitable for imaging bacteria and bacterial biofilms. The spectral resolution is on the order of 100 meV, which in combination with their intrinsic spectral properties is sufficient to provide good differentiation of classes of biomolecules (26, 43; X. Zhang, T. Araki, A. P. Hitchcock, J. R. Lawrence, and G. G. Leppard, unpublished data). Through the application of tunable soft X rays and appropriate analysis of X-ray absorption spectra in the form of NEXAFS image sequences (15), quantitative chemical mapping at a spatial scale below 50 nm may be achieved. With use of the appropriate spectral range, NEXAFS microscopy provides detailed, quantitative speciation and elemental analysis with parts-per-thousand local and parts-per-million global sensitivities with transmission detection. Soft X-ray microscopy provides a combination of suitable spatial resolution and chemical information at a microscale. In addition, soft X rays interact with nearly all elements and also allow mapping of chemical species based on bonding structure (2). Further, the method uses the intrinsic X-ray absorption properties of the sample, thus eliminating the need for addition of reflective, absorptive, or fluorescent probes and markers that may introduce artifacts or complicate interpretation. It is...

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Section: Resultssupporting

confidence: 82%

Scanning Transmission X-Ray, Laser Scanning, and Transmission Electron Microscopy Mapping of the Exopolymeric Matrix of Microbial Biofilms

Lawrence¹,

Swerhone²,

Leppard³

et al. 2003

Appl Environ Microbiol

Self Cite

327

270

View full text Add to dashboard Cite

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“…Application of synchrotron-based spectromicroscopy to study the structure of biomaterials (Hitchcock et al, 2002) now affords the opportunity to adapt novel techniques to investigate nanoscale processes in C chemistry and structural assembly of soil aggregates. Recent attempts by Schmidt et al (2003) show that spectromicroscopy can be used to capture the microscale variability of SOM in hydrated soil samples dispersed in aqueous media.…”

Section: Resolving Key Questions In Soil Biogeochemistrymentioning

confidence: 99%

Organic Carbon Chemistry in Soils Observed by Synchrotron-Based Spectroscopy

Lehmann

Solomon

2010

Developments in Soil Science

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“…This result was found for samples investigated dry, rehydrated with deionized water, and even under a buffer solution of 0.01 mg mL −1 fibrinogen. [28] Spun-cast thin films of blends of polystyrene (PS) and polymethylmethacrylate (PMMA) are being used as model substrates for both X-PEEM and STXM studies of preferential protein attachment. The morphology and surface chemistry of the PS/PMMA substrate has been investigated extensively by STXM, X-PEEM, and AFM.…”

Section: Biomaterials-stxm Versus X-peem To Study Protein Adsorption mentioning

confidence: 99%

“…[27] Biomaterials optimization. [28,29] All of the results in this article have been the subject of other publications as cited, although additional results are presented in each case. …”

Section: Introductionmentioning

confidence: 99%

Chemical Mapping of Polymer Microstructure Using Soft X‐Ray Spectromicroscopy

et al. 2005

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Recently, synchrotron-based soft X-ray spectromicroscopy techniques have been applied to studies of polymer microstructure at the ∼50 nm spatial scale. Functional group based chemical speciation and quantitative mapping is provided by near edge X-ray absorption fine structure spectral (NEXAFS) contrast. The techniques, sample data, and analysis methods of scanning transmission X-ray microscopy (STXM) and X-ray photoemission electron microscopy (X-PEEM) are outlined. The capabilities of STXM are illustrated by results from recent studies of (a) controlled release microcapsules and microspheres, (b) microcapsules being developed for gene therapy applications, (c) conducting polymer films studied in the presence of electrolyte and under potential control, and (d) studies of protein interactions with patterned polymer surfaces. In the latter area, the capabilities of STXM and X-PEEM are compared directly.

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Towards practical soft X-ray spectromicroscopy of biomaterials

Cited by 73 publications

References 26 publications

Scanning Transmission X-Ray, Laser Scanning, and Transmission Electron Microscopy Mapping of the Exopolymeric Matrix of Microbial Biofilms

Scanning Transmission X-Ray, Laser Scanning, and Transmission Electron Microscopy Mapping of the Exopolymeric Matrix of Microbial Biofilms

Organic Carbon Chemistry in Soils Observed by Synchrotron-Based Spectroscopy

Chemical Mapping of Polymer Microstructure Using Soft X‐Ray Spectromicroscopy

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