Standards for the application of X-ray microanalysis to biological specimens

A review on the subject of compounds used as standards for biological X-ray microanalysis is presented. The general approach used for standardization has been to use standards which resemble the specimen closely in composition. Thus, standards based on proteins have been used for analysis of quench-frozen cryosectioned specimens, whereas standards based on embedding resins have been used for resin-embedded material. The properties of, and problems associated with, each type of standard are recognized and have been well documented. The choice and analysis of standard should not be a drawback to fully quantitative analysis of biological material. Attention is drawn to the fact that the problems associated with any quantification procedure need to be kept in mind when analysis of standards is undertaken.     

X-ray Absorption Spectroscopy and Atomic Force Microscopy of Films Generated from Organosulfur Extreme-Pressure (EP) Oil Additives

This study examines the interaction of sulfur-based oil additives on steel. Sulfurized isobutylene, dialkyldithiocarbamate and sulfurized esters were the additives investigated in this report. For the first time, X-ray absorption near edge structure (XANES) spectroscopy has provided detailed insight into the chemistry of both the thermo-oxidative and tribochemical films generated from these additives. It was found that the chemical nature of these films was strongly dependent on the operating environment for the additives. The XANES revealed that thermally, all three S additives reacted very similarly with steel to form a film mainly comprised of iron sulfate at temperatures as low as 100 °C. The ample supply of diffused oxygen from the base oil along with oxide naturally present on the substrate allowed for the complete oxidation of the S from the decomposed additive to iron sulfate. Tribochemical films were comprised of different forms of sulfur than observed for the thermo-oxidative films. The moderate AW conditions yielded a mixture of both oxidized and reduced forms of sulfur, with pyrite, FeS₂, being the major constituent. Rubbing between the steel pin and the substrate partially depleted the oxide layer present, allowing the additive to interact intimately with the fresh substrate, yielding FeS₂. Under extreme-pressure conditions, complete removal of the oxide layer occurred with a drastic increase in the interfacial temperatures between the pin and v-block, allowing for the complete thermal decomposition of FeS₂ to FeS to occur. AFM imaging of the AW films revealed the presence of tiny smooth domains randomly oriented, which were completely different from the pad-like structures observed for AW films generated from a typical ashless thiophosphate additive. The inability of the sulfur-based additives to form large pad-like structures, which can ultimately support the load, resulted in poor AW protection to the metal.     

A new sample preparation method for biological soft X-ray microscopy: nitrogen-based contrast and radiation tolerance properties of glycol methacrylate-embedded and sectioned tissue

We describe the preparation of a biological tissue for imaging in a transmission soft X-ray microscope. Sections of exocrine pancreas embedded in glycol methacrylate polymer, an embedding medium widely used in visible light and electron microscopy, were examined. Contrast was based primarily on the nitrogen content of the tissue, and dual-wavelength imaging at the nitrogen K-shell absorption edge was used to map the distribution and provide quantitative densitometry of both the protein and embedding matrix components of the sample. The measurements were calibrated by obtaining the absorption spectrum of protein near the nitrogen edge. The contrast was consistent and reproducible, making possible the first large-scale X-ray microscopic study on sections of plastic-embedded soft tissue. At radiation doses of up to 10⁸ Gray, much more than required for routine imaging, no distortion and little mass loss were observed. This sample preparation method should permit routine imaging of tissues in X-ray microscopes, previously a difficult task, as well as multimodal imaging (using visible light, X-ray, electron, and scanned probe microscopies) on the same sample.     

Synchrotron radiation applied to the microscopical examination of solid state and biological materials in the X-ray region

The paper describes the properties of synchrotron radiation and makes some comparison with other X-ray sources. The use of synchrotron radiation for various types of X-ray imaging is discussed with reference to specific examples drawn from work carried out recently at the SERC Daresbury Laboratory.     

熔融制样-波长色散X射线荧光光谱法测定红土镍矿中铁、镍、硅、铝、镁、钙、钛、锰、铜和磷

建立了熔融制样-波长色散X射线荧光光谱法测定红土镍矿中铁、镍、硅、铝、镁、钙、钛、锰、铜和磷含量的方法.采用经1000℃灼烧后的铁矿标准样品为基体,添加相关待测元素的高纯氧化物和标准溶液制作校准曲线用的校准样品,确定了助熔剂、熔融时间、稀释比、脱模剂和基体效应校正方式等试验条件.样品分析结果进行烧失量校正,与湿法分析结果的相对标准偏差介于0.219％～2.817％之间,满足红土镍矿检测需要.     

Characterization of the effective atomic number for first row transition elements by the ratio of coherent to compton scattering intensities obtained by wavelength dispersive X-ray fluorescence

The effective atomic numbers of compounds of the first row transition elements were determined experimentally by a scattering method using wavelength dispersive X-ray fluorescence spectrometer. A calibration curve was created by using the intensity ratios of coherent to Compton scattered peaks of pure elements from atomic number 13–48. This relationship was employed to determine the effective atomic numbers of the compounds. The effective atomic numbers were also calculated by using empirical formulas from the literature. Mass attenuation coefficients were calculated using software. The experimentally measured values of the effective atomic numbers with the calculated values by empirical formulas were comparable.     

The application of X-ray analysis in the transmission electron analytical microscope (T.E.A.M.) to the quantitative bulk analysis of biological microsamples

This paper describes: (a) methods for the high and low temperature ashing of biological tissue samples about 10^?4–10^?5 g in weight; (b) a method of spraying the solubilized ash on to specimen grids which produces droplets of a size convenient for their quantitative analysis in a transmission electron analytical microscope (T.E.A.M.), but without the need to know the actual volume of individual droplets; (c) the construction of standard curves for determining the absolute concentration of Na, Mg, P, S, K, and Ca simultaneously in a given sample. For most of the elements the error in analysis was about 5% or less. It was proposed that the technique is useful for detecting local element-compartments in complex tissues, and thus may be an important aid in the interpretation of the results of point analyses performed on sections taken from adjacent micro-volumes.     

The measurement of blood flow waveforms from X-ray angiography. Part 2: Application to video recordings of digital subtraction angiography.

A method of measuring blood flow from X-ray angiograms recorded on cine film, by obtaining contrast mass values at numerous positions along individual vessels and at multiple instants of time, has been previously reported. In the present work it was hypothesized that the signal-to-noise limitations of recordings on video tape could be overcome by recording already-subtracted angiograms from a digital subtraction angiographic (DSA) system and that the spatial resolution of video was adequate to use a similar measurement method. Validation experiments were recorded, in which flows measured using a calibrated electromagnetic flowmeter passed through tubes of 4-7 mm diameter, during injections of 2-5 ml contrast medium. The video sequences were computer analysed and produced angiographic flow measurements agreeing with the electromagnetic flows to within 5 per cent. A case study of a clinical carotid artery DSA in a patient with secondary carcinoma metastases in the brain is described. It is concluded that accurate flow measurements can be made from DSA video recordings.     

Determination of the physical structure of biological materials at biosensor interfaces by techniques of increasing magnification from microscopic to molecular scale

Chemical selectivity of biosensors is derived from biological materials interfaced to the surface of transducing devices. Molecular recognition events lead to macroscopic function suitable for analytical measurements. The structure-function relationships of biochemical species at interfaces must be established to characterize and optimize biosensor operation. The techniques of ellipsometry, fluorescence microscopy, electron microscopy, and scanning tunneling microscopy are used to investigate the structure of monolayers and multilayers of proteins and lipids at interfaces that are prepared by Langmuir-Blodgett techniques and by self-assembly from bulk solution. The relative merits and limitations of the measurement techniques in the determination of aspects of interfacial structure are considered.