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At least 19 recordsLinked to original sources
PubMed2004

Detection of phosphorylated peptides in proteomic analyses using microfluidic compact disk technology.

A compact disk (CD)-based microfluidic method for selective detection of phosphopeptides by mass spectrometry is described. It combines immobilized metal affinity chromatography (IMAC) and enzymatic dephosphorylation. Phosphoproteins are digested with trypsin and processed on the CD using nanoliter scale IMAC with and without subsequent in situ alkaline phosphatase treatment. This is followed by on-CD matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. Dephosphorylation of the IMAC-enriched peptides allows selective phosphopeptide detection based on the differential mass maps generated (mass shifts of 80 Da or multiples of 80 Da). The CD contains 96 microstructures, each with a 16 nL IMAC microfluidic column. Movement of liquid is controlled by differential spinning of the disk. Up to 48 samples are distributed onto the CD in two equal sets. One set is for phosphopeptide enrichment only, the other for identical phosphopeptide enrichment but combined with in situ dephosphorylation. Peptides are eluted from the columns directly into MALDI target areas, still on the CD, using a solvent containing the MALDI matrix. After crystallization, the CD is inserted into a MALDI mass spectrometer for analysis down to the femtomole level. The average success rate in phosphopeptide detection is over 90%. Applied to noncharacterized samples, the method identified two novel phosphorylation sites, Thr 735 and Ser 737, in the ligand-binding domain of the human mineralocorticoid receptor.

Alkaline Phosphatase↗
PubMed2003

The compact disk as radon detector--a laboratory study of the method.

The radon absorption ability and the track etch properties of the polycarbonate material of commercial compact disks make them very useful as sensitive retrospective 222Rn detectors. The basic idea is to remove, after exposure, a surface layer that is thicker than the range of the alpha particles of the 222Rn and 220Rn progenies and to count the electrochemically etched tracks at the corresponding depths (>80 microm). The effects on the response due to differences in pressure, temperature, and humidity have been studied experimentally. The effect of the growing of 210Po after long-term exposures was also estimated. The effect of all listed factors except the temperature is either absent or restricted to maximum--about 10% for the very extreme cases. The variation of the response at 83 microm depth over the temperature interval 15-25 degrees C is +/-12% around the 20 degrees C value. The dependence of the calibration factor on the etched depth beneath the surface was studied at 4 different temperatures within the range expected indoors. The results show that the depth dependence is exponential with the parameters of the exponent also being dependent on the temperature. In practice, using the track density obtained in two or more depths beneath the compact disk's front surface, an a posteriori temperature correction could be made. By this correction it is possible to substantially reduce the bias in the results due to the unknown temperature during exposure. The results imply that by using home stored compact disks long-term retrospective 222Rn measurements could be made with an uncertainty that could be potentially better than 10%. The useful range of the method starts at about 3 Bq m(-3) (for 10 y exposure time) and appears to cover practically the whole range of indoors 222Rn concentrations.

Alpha Particles↗
PubMed1993

Auditory demonstrations on compact disk for large N.

The popular compact disk entitled Auditory Demonstrations, sponsored by the Acoustical Society of America, includes a number of demonstrations that lead to quantitative results. Those demonstrations are evaluated here in the context of a sizeable class in a lecture room. Demonstrations concern masking, loudness, and pitch; specifically they are numbers 2, 3, 6, 7, 8, 9, 10, 11, 12, 14, 15, 16, 17, 20, and 25. The evaluations find that most of the demonstrations successfully illustrate psychoacoustical principles in a classroom context; others are less successful or require special circumstances for success. Explanations for success and failure are offered, together with some suggestions for optimizing the chances of success.

Acoustic Stimulation↗
PubMed2001

Fungal bioturbation paths in a compact disk.

We report here on bioturbation traces, with micro-dendrite textures, composed of a mixture of altered aluminum and polycarbonate, which have been developed in a common compact disk (CD), destroying information pits. Fungal hyphae proliferated in these deteriorated zones, and Geotrichum-type fungus was isolated from surface-sterilized CD fragments. The severe biodeterioration described is attributed to the slow growth of this arthroconidial fungus on the CD material in the tropical indoor environment of Belize, Central America (approximately 30 degrees C, approximately 90% humidity).

Aluminum↗
PubMed2006

Microfluidic analysis of antibody specificity in a compact disk format.

A new and flexible technology for high throughput analysis of antibody specificity and affinity is presented. The method is based on microfluidics and takes advantage of compact disks (CDs) in which the centrifugal force moves fluids through microstructures containing immobilized metal affinity chromatography columns. Analyses are performed as a sandwich assay, where antigen is captured to the column via a genetically attached His6-tag. The antibodies to be analyzed are applied onto the columns. Thereafter, fluorescently labeled secondary antibodies recognize the bound primary antibodies, and detection is carried out by laser-induced fluorescence. The CDs contain 104 microstructures enabling analysis of antibodies against more than 100 different proteins using a single CD. Importantly, through the three-dimensional visualization of the binding patterns in a column it is possible to separate high affinity from low affinity binding. The method presented here is shown to be very sensitive, flexible and reproducible.

Animals↗
PubMed2002

Cell analysis system based on compact disk technology.

BACKGROUND: A cell analysis system was developed to enumerate and differentiate magnetically aligned cells selected from whole blood. The cellular information extracted is similar to the readout of musical information from a compact disk (CD). Here we describe the optical design and data processing of the system. The performance of the system is demonstrated using fluorescent-labeled cells and beads. Materials and Methods System performance was demonstrated with 6-microm polystyrene beads labeled with magnetic nanoparticles and allophycocyanin (APC) and immunomagnetically aligned leukocytes, fluorescently labeled with Oxazine750 and CD4-APC, CD8-Cy5.5, and CD14-APC/Cy7 in whole blood. RESULTS: The sensitivity of the system was demonstrated using APC-labeled beads. With this system, beads containing 333 APC molecules could easily be resolved from the background. This level of sensitivity was not achievable with a commercial flow cytometer. A maximum of 20,000 immunomagnetically labeled cells could be aligned and analyzed in between 0.6 m of Ni lines, distributed over a surface area of 18 mm(2) and extracted from a blood volume that depended on the height of the chamber. The utility of the system was demonstrated by performing a three-color CD4-CD8-CD14 assay. CONCLUSIONS: We built a cell analysis system based on immunomagnetic cell selection and alignment and analysis of fluorescent signals employing CD-technology that is as good or better than current commercial analyzers. The cell analysis can be performed in whole blood or any other type of cell suspension without extensive sample preparation.

Animals↗
PubMedDate not supplied

["Tactic and technique" for keeping medical information on compact disk].

Based on experience accumulated in time of realization and publication the Atlas of Surgical Pathology, the work present the main methodologies elements used for keeping of some elements so called by "tactics" (cases, following in time) it's insist on photographic technology in medical practice with special reference to digital technology, later on being also described the elements of processing and keeping the medical information on compact disk. Finally are enunciated a series of virtual programs with surgical specific proposed for be created and maintained on-line, with the support of all interested surgeons.

Atlases as Topic↗
PubMed2001

Cell analysis system based on immunomagnetic cell selection and alignment followed by immunofluorescent analysis using compact disk technologies.

BACKGROUND: Although the flow cytometer has become the standard in cell analysis, it has limitations. Recently, we introduced a new cell analysis method based on immunomagnetic selection and aligning of cells. No flow system is needed and cell analysis can be performed in whole blood. METHODS: Whole blood is incubated with fluorescent labels and immunomagnetic nanoparticles. The blood is injected into a capillary that is in a strong magnetic field. The immunomagnetic-labeled cells move upward and align themselves along ferromagnetic lines present on the upper surface of the capillary. An optical focus and tracking system analogous to that used in a conventional compact disk player focuses a 635-nm laser-diode on the magnetically aligned cells. The emitted fluorescence signals are projected on two photomultipliers. Allophycocyanin (APC)-labeled CD4 (CD4-APC) and Cyanin5.5 (Cy5.5)-labeled CD8 (CD8-Cy5.5) antibodies and Oxazine750, all red excited, are used as fluorescent labels. RESULTS: A differential white blood cell count performed in whole blood is obtained using the CD4-APC in combination with Oxazine750. The results are compared with the Technicon-H1 hematology analyzer. Correlation coefficients of 0.91 for neutrophilic granulocytes, 0.93 for lymphocytes, 0.93 for monocytes, and 0.96 for eosinophilic granulocytes were obtained. Immunofluorescence is demonstrated using CD4-APC and CD8-Cy5.5. The absolute counts obtained for CD4+ and CD8+ are compared with the Coulter Epics XL flow cytometer. Correlation coefficients of, respectively, 0.91 and 0.94 were obtained. CONCLUSION: We conclude that our system is as capable as a standard flow cytometer or hematology analyzer for a reliable routine white blood cell analysis, including immunophenotyping, and can be used as an easy-to-handle disposable white blood cell test.

CD4 Antigens↗
PubMed2004

Integrated sample preparation and MALDI mass spectrometry on a microfluidic compact disk.

High-throughput microfluidic processing of protein digests integrated with matrix-assisted laser desorption/ionization (MALDI) mass spectrometry on a compact disk (CD) is described. Centrifugal force moves liquid through multiple microstructures, each containing a 10-nL reversed-phase chromatography column. The CD enables parallel preparation of 96 samples with volumes ranging from one to several microliters. The peptides in the digests are concentrated, desalted, and subsequently eluted from the columns directly into MALDI target areas (200 x 400 microm) on the CD using a solvent containing the MALDI matrix. After crystallization, the CD is inserted into the MALDI instrument for peptide mass fingerprinting and database identification at a routine sensitivity down to the 200-amol level. Detection of proteolytic peptides down to the 50-amol level is demonstrated. The success rate of the CD technology in protein identification is about twice that of the C(18) ZipTips and standard MALDI steel targets. The CDs are operated using robotics to transfer samples and reagents from microcontainers to the processing inlets on the disposable CD and spinning to control the movement of liquid through the microstructures.

Journal Article↗
PubMed2004

Theoretical and experimental study of the surface plasmon resonance effect on a recordable compact disk.

The surface plasmon resonance (SPR) effect on noble-metal surfaces has been explored by several investigators for the development of chemical and biological sensors as well as for the design of optical devices for other applications. The effect can be observed by use of prism couplers, diffraction gratings, and specially configured optical fibers. In an attempt to seek a new configuration that minimizes costs of fabricating media that support SPR in conjunction with designing a new format suitable for large-scale chemical and biological sensing, I have investigated the feasibility of using a commercially available, gold-type, recordable compact disk for observation of the SPR phenomenon. Experimental and theoretical results of this investigation are reported.

Journal Article↗
PubMed1994

[Comparative speech audiometry examinations with compact disk and cassette tape].

We compared the results of speech audiometry, performed with recorded tape and compact disc recordings (CD), respectively, in 21 subjects with bone-conduction hearing losses CHL and 27 subjects with sensorineural hearing losses. In all cases, speech audiometry was performed by using the 1969 Freiburg speech test material based on the German DIN standard 45626. In subjects with air-bone gaps, the hearing loss for speech was 2.3 dB smaller if tested by use of CD and speech recognition was 10.5 points better. Smaller differences could be found in sensorineural HL. For CD, hearing losses were 1.2 dB smaller and speech recognition scores were increased by only 1 point. These differences were not statistically significant. These findings show that calculations of hearing impairments and individual hearing handicaps from tables commonly used in Germany in medicolegal expert evaluations should not be changed if CD records are used that are based on 1969 DIN 4526 recordings. However, the handling of compact discs has many advantages in relation to the handling of tapes.

Adult↗
PubMed1992

[Comparative speech audiometry studies with compact disk].

We compared the results of speech audiometry, performed with recorded tape and CD, respectively, in 21 subjects with bone conduction hearing loss, and in 27 subjects with sensorineural hearing loss. In all cases, speech audiometry was performed by using the German Freiburg speech test material, recorded in 1969 on the basis of the German DIN standard 45626. In subjects with an air-bone gap, the hearing loss for speech is 2.3 dB smaller if tested by use of CD, and the speech recognition is 10.5 points better, respectively. Smaller differences could be found in sensorineural HL: For CD, the hearing loss is 1.2 dB smaller, and the speech recognition score is raised by only 1 point. These differences are statistically not significant, and the calculation of hearing impairment and individual hearing handicap from the tables commonly used in Germany in medicolegal expert opinions must not be corrected, if records on CD are used basing on the recordings from 1969 according to DIN 45626. The handling of compact discs has many advantages in relation to the handling of tapes.

Adult↗
PubMed2004

Whole-cell-reporter-gene-based biosensing systems on a compact disk microfluidics platform.

Biosensing systems such as reporter-gene-based whole-cell assays are increasingly finding applications in biological and environmental screening. A whole-cell approach to such analyses can provide valuable information about the bioavailable level of a compound of interest. These biosensing systems rely on the molecular recognition of a specific analyte by a regulatory protein and, therefore, can detect low levels of the target analyte. In this study, Escherichia coli cells containing plasmid pSD10 were engineered to sense the model target analytes arsenite and antimonite, the target analytes in this study. The biosensing system takes advantage of the recognition of the regulatory protein, ArsR, for arsenite and antimonite to produce the reporter protein, which in this case is GFPuv. The fluorescence emitted by the GFPuv in the cells can be directly related to the concentration of the analyte in the cell, making this biosensing system useful in the detection of arsenite and/or antimonite in a variety of samples. Miniaturization of biosensing systems can further enhance their utility by decreasing reagent consumption and analysis time and by allowing for the high-throughput screening of samples. To that end, we employed a microcentrifugal microfluidics platform that has low power, space, and reagent requirements, increased speed of detection, and the potential for portability. Herein, we demonstrate for the first time the adaptation of a whole-cell sensing system to a microcentrifugal microfluidics platform. Moreover, we were able to detect our target analytes in a rapid and sensitive manner compared to conventional sensing methods.

Antimony↗
PubMed1998

[The hemodynamics laboratory without cine film. A comparison between the production costs of 35-mm film and the recordable compact disk].

BACKGROUND: For more than 35 years, cine film has been used as the standard recording medium for coronary angiography. At present, the DICOM and the CD-R format have been established as the standard media for the transport of cardiac angiographic images in place of cine angiographic film. The aim of the study was to compare the media production costs between two cardiac catheterization laboratories, with and without cine film. METHODS: We collected data from two different laboratories in the same geographic area which perform about 1000 procedures/year, using a similar digital x-ray imaging system. In one lab, images are recorded on 35-mm film at 25 frames/sec. In the other one, the image support is based on a CD-R. For each laboratory we considered both direct patient and variable equipment costs. Direct patient costs in the film-lab include: cine film, processing chemicals, processing labor, chemical disposal, maintenance; in the digital lab: CD-R costs and masterization time. Equipment costs in the film-lab include: cine camera, cine film processor and cine projector; in the filmless lab a DICOM formatter and a review workstation. The equipment amortization costs are considered over a three-year period. RESULTS: Total direct patient costs are 90,000 lira for the film and 14,000 lira for the CD-R. Equipment costs are 193,000,000 lira in the film-lab and 150,000,000 lira in the filmless one. Overall cost per patient is 154,300 lira for the cine film and 64,000 lira for the CD-R. CONCLUSIONS: This study shows that the media costs per patient for a digital DICOM CD-R format system are substantially less than for 35-mm film, permitting savings of more than 90,000,000 Italian lira per year in a mid-volume cardiac catheterization laboratory.

Compact Disks↗
PubMed1992

Multianalyte microspot immunoassay. The microanalytical 'compact disk' of the future.

Throughout the 1970s, controversy centered both on immunoassay 'sensitivity' per se and on the relative sensitivities of labelled antibody and labelled analyte methods. Our own theoretical studies in this period revealed that radioimmunoassay (RIA) sensitivities could be surpassed only by the use of very high specific activity non-isotopic labels in 'non-competitive' designs, preferably based on the use of monoclonal antibodies. The time-resolved fluorescence methodology known as Delfia - developed in collaboration with the instrument manufacturer LKB/Wallac - represented the first commercial 'ultra-sensitive' non-isotopic technique based on these theoretical insights, the same concepts being subsequently adopted in comparable methodologies relying on the use of chemiluminescent and enzyme labels. However, a second advantage of high specific activity labels is that they permit the development of 'multi-analyte' immunoassay systems combining ultra-sensitivity with the simultaneous measurement of tens, hundreds or thousands of analytes in a small biological sample. This possibility relies on simple, albeit hitherto unexploited, physicochemical concepts. The first is that all immunoassays rely on measurement of Ab occupancy by analyte. The second is that, provided the Ab concentration used is 'vanishingly small', fractional Ab occupancy is independent of both Ab concentration and sample volume. This leads to the notion of 'ratiometric' immunoassay, involving measurement of the ratio of signals (eg fluorescent signals) emitted by two labelled Ab's, the first ('sensor' Ab) deposited as a microspot on a solid support, the second a 'developing' Ab directed against either occupied or unoccupied sensor Ab binding sites.(ABSTRACT TRUNCATED AT 250 WORDS)

Binding, Competitive↗
PubMed1991

Multianalyte microspot immunoassay--microanalytical "compact disk" of the future.

Throughout the 1970s, controversy centered both on immunoassay "sensitivity" per se and on the relative sensitivities of labeled antibody (Ab) and labeled analyte methods. Our theoretical studies revealed that RIA sensitivities could be surpassed only by the use of very high-specificity nonisotopic labels in "noncompetitive" designs, preferably with monoclonal antibodies. The time-resolved fluorescence methodology known as DELFIA--developed in collaboration with LKB/Wallac--represented the first commercial "ultrasensitive" nonisotopic technique based on these theoretical insights, the same concepts being subsequently adopted in comparable methodologies relying on the use of chemiluminescent and enzyme labels. However, high-specific-activity labels also permit the development of "multianalyte" immunoassay systems combining ultrasensitivity with the simultaneous measurement of tens, hundreds, or thousands of analytes in a small biological sample. This possibility relies on simple, albeit hitherto-unexploited, physicochemical concepts. The first is that all immunoassays rely on the measurement of Ab occupancy by analyte. The second is that, provided the Ab concentration used is "vanishingly small," fractional Ab occupancy is independent of both Ab concentration and sample volume. This leads to the notion of "ratiometric" immunoassay, involving measurement of the ratio of signals (e.g., fluorescent signals) emitted by two labeled Abs, the first (a "sensor" Ab) deposited as a microspot on a solid support, the second (a "developing" Ab) directed against either occupied or unoccupied binding sites of the sensor Ab. Our preliminary studies of this approach have relied on a dual-channel scanning-laser confocal microscope, permitting microspots of area 100 microns 2 or less to be analyzed, and implying that an array of 10(6) Ab-containing microspots, each directed against a different analyte, could, in principle, be accommodated on an area of 1 cm2. Although measurement of such analyte numbers is unlikely ever to be required, the ability to analyze biological fluids for a wide spectrum of analystes is likely to transform immunodiagnostics in the next decade.

Binding, Competitive↗