Development of Micro-scale Sample Preparation and Prefractionation Methods in LC-MS-Based Proteomic Studies

Abstract A new automated method of sample preparation and introduction that may circumvent the problems of measurement of micro-scale quantities of samples and preparation of serum or soluble blood fraction is being developed for use in fast analytical systems based on the GeMSAEC principle. This method involves the dynamic introduction of reagents or blood samples into the centrifugal field of the Fast Analyzer during rotation of the rotor. The rotor can be designed to fractionate the fluid into discrete, measured portions that are transferred to the cuvets for photometric monitoring. Whole-blood samples can be directly introduced into the rotating rotor where, after fractionation into measured volumes, the particulates are automatically removed by the centrifugal field and the resulting measured quantities of plasma are transferred directly to the cuvets. The dynamic proportionation of reagents, serum, and whole blood has been shown to be precise, and automatic analyses for specific chemicals have been performed on whole blood samples with a coefficient of variation of less than 2%.

Download Full-text

Microstructural Origin of Superior Compressive Ductility of a Nanocrystalline Metal

Materials Science Forum ◽

10.4028/www.scientific.net/msf.633-634.73 ◽

2009 ◽

Vol 633-634 ◽

pp. 73-84

Author(s):

Deng Pan ◽

S. Kuwano ◽

T. Fujita ◽

M. W. Chen

Keyword(s):

Sample Preparation ◽

Grain Boundary ◽

Grain Growth ◽

Room Temperature ◽

Focused Ion Beam ◽

Ion Beam ◽

Ex Situ ◽

Preparation Technique ◽

Micro Scale ◽

Sample Preparation Technique

Ultra-large compressive plasticity at room temperature has recently been observed in electrodeposited nanocrystalline nickel (nc-Ni) under micro-scale compression (Pan, Kuwano, Fujita and Chen: Nano Lett. Vol. 7 (2007), p. 2108). With aid of a TEM sample preparation technique employing focused ion beam (FIB), TEM observations on deformed nc-Ni evidenced deformation-induced microstructural evolution of nc-Ni at a variety of strain levels: Whilst the deformation increases, substantial grain growth is uncovered in the nc-Ni. No apparent ex situ evidence of intragranular dislocation activities is found in the deformed sample. As thermal diffusion plays an insignificant role in the deformation in nc-Ni at room temperature (~0.17Tm), this premium plasticity is achieved in accommodation with the grain-boundary-mediated deformation, with assistance of extensive grain growth that is mainly driven by high stresses at steady plastic flow.

Download Full-text

Macro- and Micro-Scale Effects of Pluviation Based Sample Preparation in DEM

GeoCongress 2012 ◽

10.1061/9780784412121.245 ◽

2012 ◽

Cited By ~ 1

Author(s):

M. L. Bernhardt ◽

C. O'Sullivan ◽

G. Biscontin

Keyword(s):

Sample Preparation ◽

Scale Effects ◽

Micro Scale

Download Full-text

Pollutant Identification by Selected Area Electron Diffraction

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100052675 ◽

1974 ◽

Vol 32 ◽

pp. 532-533

Author(s):

R. E. Ferrell ◽

G. G. Paulson ◽

C. W. Walker

Keyword(s):

Thermal Treatment ◽

Electron Diffraction ◽

Sample Preparation ◽

Crystal Structures ◽

Water Samples ◽

Environmental Samples ◽

Short Review ◽

Selected Area Electron Diffraction ◽

Multiple Component

Selected area electron diffraction (SAD) has been used successfully to determine crystal structures, identify traces of minerals in rocks, and characterize the phases formed during thermal treatment of micron-sized particles. There is an increased interest in the method because it has the potential capability of identifying micron-sized pollutants in air and water samples. This paper is a short review of the theory behind SAD and a discussion of the sample preparation employed for the analysis of multiple component environmental samples.

Download Full-text

Preparation of Thin Cadmium Telluride Samples for Electron Microscope Studies

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100052754 ◽

1974 ◽

Vol 32 ◽

pp. 548-549

Author(s):

T. J. Magee ◽

J. Peng ◽

J. Bean

Keyword(s):

Electron Microscope ◽

Sulfuric Acid ◽

Solid State ◽

Sample Preparation ◽

Cadmium Telluride ◽

Potassium Dichromate ◽

Optical Components ◽

The Past ◽

Solid State Devices

Cadmium telluride has become increasingly important in a number of technological applications, particularly in the area of laser-optical components and solid state devices, Microstructural characterizations of the material have in the past been somewhat limited because of the lack of suitable sample preparation and thinning techniques. Utilizing a modified jet thinning apparatus and a potassium dichromate-sulfuric acid thinning solution, a procedure has now been developed for obtaining thin contamination-free samples for TEM examination.

Download Full-text

A Freeze Shadow Technique for the Preparation of Soft Paint Latexes for Electron Microscopy

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100079097 ◽

1977 ◽

Vol 35 ◽

pp. 314-315

Author(s):

Earl R. Walter ◽

Glen H. Bryant

Keyword(s):

Sample Preparation ◽

High Vacuum ◽

Vacuum System ◽

Latex Particles ◽

New Methods ◽

Diffusion Pump ◽

Film Forming ◽

Routine Basis ◽

Distribution Studies ◽

Preparation Techniques

With the development of soft, film forming latexes for use in paints and other coatings applications, it became desirable to develop new methods of sample preparation for latex particle size distribution studies with the electron microscope. Conventional latex sample preparation techniques were inadequate due to the pronounced tendency of these new soft latex particles to distort, flatten and fuse on the substrate when they dried. In order to avoid these complications and obtain electron micrographs of undistorted latex particles of soft resins, a freeze-dry, cold shadowing technique was developed. The method has now been used in our laboratory on a routine basis for several years.The cold shadowing is done in a specially constructed vacuum system, having a conventional mechanical fore pump and oil diffusion pump supplying vacuum. The system incorporates bellows type high vacuum valves to permit a prepump cycle and opening of the shadowing chamber without shutting down the oil diffusion pump. A baffeled sorption trap isolates the shadowing chamber from the pumps.

Download Full-text

SEM evaluation of the TEM cold-stage double-film specimen

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100111471 ◽

1984 ◽

Vol 42 ◽

pp. 272-273

Author(s):

Jayesh Bellare

Keyword(s):

Spatial Distribution ◽

Sample Preparation ◽

Sample Thickness ◽

Specimen Preparation ◽

Preparation Technique ◽

Liquid Sample ◽

Thin Specimen ◽

Sample Preparation Technique ◽

Cold Stage ◽

Film Specimen

Seeing is believing, but only after the sample preparation technique has received a systematic study and a full record is made of the treatment the sample gets.For microstructured liquids and suspensions, fast-freeze thermal fixation and cold-stage microscopy is perhaps the least artifact-laden technique. In the double-film specimen preparation technique, a layer of liquid sample is trapped between 100- and 400-mesh polymer (polyimide, PI) coated grids. Blotting against filter paper drains excess liquid and provides a thin specimen, which is fast-frozen by plunging into liquid nitrogen. This frozen sandwich (Fig. 1) is mounted in a cooling holder and viewed in TEM.Though extremely promising for visualization of liquid microstructures, this double-film technique suffers from a) ireproducibility and nonuniformity of sample thickness, b) low yield of imageable grid squares and c) nonuniform spatial distribution of particulates, which results in fewer being imaged.

Download Full-text