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PubMed2004

An evaluation of the utility of performing body fluid counts on the coulter LH 750.

Performing a manual body fluid count is a tedious, time-consuming, and frequently imprecise process for any clinical laboratory. The ability to perform many of these counts with an automated hematology analyzer has the real potential of making a major impact on laboratory precision and productivity. The manual chamber count is fraught with many variables and is often very technologist/technique dependent, often leading to inaccuracies in test results. Our laboratory undertook a series of studies designed to evaluate the capability of the Coulter LH 750 hematology analyzer to supplement our current manual method in the performance of body fluid analysis. First, we established the performance of our current manual counting method, the gold standard against which LH 750 performance would be judged. We looked at the precision of manual cell counting by having 4 technologists perform manual counts on each of 35 spinal, synovial, peritoneal, and other fluids with white blood cell (WBC) and/or red blood cell (RBC) counts of greater than 0.3 x 10(9) /L and 0.03 x 10(12) /L, respectively. Our results support earlier reports that the variability in counts among technologists using the manual chamber cell count methods is often very significant. After each sample was manually counted, it was analyzed on the Coulter LH 750, and the results were compared with those of our current manual method. Results showed good correlation between the manual and LH 750 methods. Next, we conducted a series of separate tests to evaluate the stability of different cellular elements (WBCs and RBCs) in each of these body fluid types. The study consisted of 2 sets of 4--3-mL samples of each fluid type--which were analyzed on the LH 750 immediately on receipt and after 1 hour, 4 hours, 8 hours, 16 hours, and 24 hours. The findings suggested varying degrees of stability that were dependent on fluid type and initial cell concentration. Finally, we looked at whether it is necessary to perform background counts before analyzing each body fluid sample and the impact of automating body fluid counts on our workload.

Autoanalysis↗
PubMed2004

Evaluation of the neutralising capacity of bactec medium for some antibiotics.

Bactec medium 9240 (Becton Dickinson, MD, USA) is a blood culture medium used routinely at Sultan Qaboos University Hospital (SQUH). The medium is said to contain substances that can neutralise antibiotics and destroy leucocytes in blood samples. In this study, the ability of the medium to neutralise the effect of some antibiotics and to destroy leucocytes is investigated. Vancomycin, amoxicillin, chloramphenicol, penicillin, gentamicin, fungizone and amikacin at various concentrations were added to separate bottles of Bactec medium 9240. Escherichia coli (NCTC 10418), Pseudomonas aeruginosa (NCTC 10662), Staphylococcus aureus (NCTC 6571) and Candida albicans (ATCC 10231), each at 1 x 10(4) colony-forming units (CFU)/mL were added separately, depending on the type of antibiotic. The blood samples were incubated at 37 degrees C for seven days under manual and automated blood culture systems. Subcultures were made from the manual system and routine laboratory procedures for detection of positive cultures were followed for the automated system. The Bactec medium was found to neutralise all antibiotics up to a concentration of 100 microg/mL by the automated method but showed some variation in results by the manual system. Leucocytes were destroyed within 24 hours.

Amikacin↗
PubMed1992

Miles Technicon H.2 automated hematology analyzer.

Automated hematology analyzers are used in all large hospitals and most commercial laboratories, as well as in most smaller hospitals and laboratories, to perform complete blood counts (including white blood cell, red blood cell, and platelet counts; hemoglobin concentration; and RBC indices) and white blood cell differential counts. Our objectives in this study are to provide user guidance for selecting, purchasing, and using an automated hematology analyzer, as well as to present an overview of the technology used in an automated five-part differential unit. Specifications for additional automated units are available in ECRI's Clinical Laboratory Product Comparison System. We evaluated the Miles Technicon H.2 unit and rated it Acceptable. The information in this Single Product Evaluation is also useful for purchasing other models; our criteria will guide users in assessing components, and our findings and discussions on some aspects of automated hematology testing are common to many available systems. We caution readers not to base purchasing decisions on our rating of the Miles unit alone, but on a thorough understanding of the issues surrounding automated hematology analyzers, which can be gained only by reading this report in its entirety. The willingness of manufacturers to cooperate in our studies and the knowledge they gain through participating lead to the development of better products. Readers should refer to the Guidance Section, "Selecting and Purchasing an Automated Hematology Analyzer," where we discuss factors such as standardization, training, human factors, manufacturer support, patient population, and special features that the laboratory must consider before obtaining any automated unit; we also provide an in-depth review of cost issues, including life-cycle cost analyses, acquisition methods and costs of hardware and supplies, and we describe the Hemacost and Hemexmpt cost worksheets for use with our PresValu and PSV Manager CAHDModel software. Readers should also review the Clinical and Technical Overview and the articles "Miles Technicon H.2 Reports" and "Developing the Evaluation Reference Method."

Blood Cell Count↗
PubMed2004

A portable automated system for trace gas sampling in the field and stable isotope analysis in the laboratory.

A computer-controllable mobile system is presented which enables the automatic collection of 33 air samples in the field and the subsequent analysis for delta13C and delta18O stable isotope ratios of a carbon-containing trace gas in the laboratory, e.g. CO2, CO or CH4. The system includes a manifold gas source input for profile sampling and an infrared gas analyzer for in situ CO2 concentration measurements. Measurements of delta13C and delta18O of all 33 samples can run unattended and take less than six hours for CO2. Laboratory tests with three gases (compressed air with different pCO2 and stable isotope compositions) showed a measurement precision of 0.03 per thousand for delta13C and 0.02 per thousand for delta18O of CO2 (standard error (SE), n = 11). A field test of our system, in which 66 air samples were collected within a 24-hour period above grassland, showed a correlation of 0.99 (r2) between the inverse of pCO2 and delta13C of CO2. Storage of samples until analysis is possible for about 1 week; this can be an important factor for sampling in remote areas. A wider range of applications in the field is open with our system, since sampling and analysis of CO and CH4 for stable isotope composition is also possible. Samples of compressed air had a measurement precision (SE, n = 33) of 0.03 per thousand for delta13C and of 0.04 per thousand for delta18O on CO and of 0.07 per thousand for delta13C on CH4. Our system should therefore further facilitate research of trace gases in the context of the carbon cycle in the field, and opens many other possible applications with carbon- and possibly non-carbon-containing trace gases.

Carbon Dioxide↗
PubMed2002

The effect of sample treatment on separation profiles of tear fluid proteins: qualitative and semi-quantitative protein determination by an automated analysis system.

PURPOSE: Qualitative and quantitative determination of tear fluid components is of increasing interest in ophthalmology. Until now, for diagnosis and course control of some diseases of the anterior parts of the eye, different methods for tear fluid protein analysis are available. Results can be obtained by polyacrylamide gel electrophoresis (PAGE), immunochemistry, and high-performance liquid chromatography (HPLC). A new method for protein separation, identification and semi-quantitative determination on a chip-based micro-fluidic technique is used for the first time to investigate tear fluids. METHODS: Normal human reflex tears were obtained by stimulation with China mint oil and collected using glass capillary tubes. A lab-on-a-chip technology (developed by Agilent Technologies, Waldbronn, Germany, in co-operation with Caliper Technologies, Mountain View, California, USA) was used for separation and semi-quantitative determination of tear proteins. Tear fluid was separated on the Agilent 2100 Bioanalyzer in combination with the Protein 200 LabChip kit and the dedicated protein assay software. Time and temperature of the incubation with sample buffer were varied, and the influence of these parameters on protein separation profiles was studied. Tear proteins were also analysed by PAGE, and the results obtained by both methods were compared. RESULTS: The different proteins of tear fluid can be separated by the Agilent 2100 Bioanalyzer method in very short time. By this method, the molecular weight as well as the concentration of proteins can be determined. Data are automatically stored in digital format and can be retrieved and shared. Results of this technology were comparable with the protein pattern obtained by PAGE. It was confirmed by both methods that, depending on incubation time of tear fluid with sample buffer and on temperature, different protein pattern can be obtained from the tears of one specimen. CONCLUSION: Tear proteins - in contrast to serum or aqueous humour proteins - are very sensitive to changes in sample buffer temperature as well as incubation time with buffer. To obtain comparable results for tear fluid proteins, the sample buffer applied and the incubation time and temperature must be observed carefully. This can be demonstrated by both the new Agilent 2100 Bioanalyzer method and PAGE. These results are of importance when comparing tear fluid protein pattern for the diagnosis and course control of dry-eye syndrome and of other diseases of the anterior part of the eye.

Aqueous Humor↗
PubMed1991

A microplate version of the SOS/umu-test for rapid detection of genotoxins and genotoxic potentials of environmental samples.

The umu-microtest is a miniaturized automated short-term test version proposed for screening of umuC-dependent mutagenic potentials of chemicals relevant to environmental pollution, river water and industrial waste water. The test is based on the SOS/umu-test and has been modified in order to allow extensive testing of environmental samples. Genetically engineered Salmonella typhimurium (TA1535/pSK1002) are incubated on a microplate rotor in a sloping position for 2 h with the test samples, followed by addition of fresh culture medium to reach a 10-fold dilution of the incubation medium. 2 h later, the activity of the beta-galactosidase, which reflects umuC induction, is determined colorimetrically. The incubation of the bacteria in the presence of the test compounds as well as the assessment of beta-galactosidase activity takes place in 96-well microplates, thus enabling simultaneous screening of large numbers of samples. Data of the genotoxic potentials are available within 8 h. Computer-controlled automation is possible by using a laboratory workstation.

Azides↗
PubMed1991

The diagnosis of acute viral hepatitis A or B by microparticle enzyme immunoassay.

The traditional approach to the diagnosis of viral hepatitis has been to collect a serum sample and to test it for the presence of hepatitis B surface antigen (HBsAg), IgM to the core of HBV (anti-HBc IgM) and IgM to HAV (anti-HAV IgM) by solid phase radio- or enzyme immunoassays. Microparticle enzyme immunoassay technology (IMx-Abbott Laboratories) has been introduced as an automated carousel system for the detection of these markers. A side-by-side, blinded comparison of IMx to current IA was performed for HBsAg on 659 specimens submitted from March to July 1990, of which 72 (10.8%) were positive by AUSRIA (Abbott RIA for HBsAg) and 2 of these were discordant in IMx. Both were near the cutoff and by confirmatory testing one was positive and the other negative. Forty three percent (25/58) of frozen stored sera tested for anti-HBc IgM, by IMx, were positive by EIA (Corzyme M). One specimen near the cutoff was negative by EIA but weakly positive by IMx. Anti-HAV IgM was found in 21.8% (46/211) of sera with 100% correlation by IMx. Thus IMx had the following percent sensitivies and specificities: HBsAg; 98.6, 99.9; anti-HBc IgM; 100, 99.9; anti-HAV IgM; 100, 100. The test set-up times for the 3 markers in the IMx were similar to the RIA and EIA. The turnover time was 45 min for a full IMx carousel compared to: AUSRIA-short incubation (4 h), or long incubation (14 h); Corzyme M-short (4.75 h) or long (20 h); anti-HAV IgM (23 h).(ABSTRACT TRUNCATED AT 250 WORDS)

Automation↗
PubMed1985

Methods for rapid transmission of microbiology reports.

Rapid microbiologic testing is of value to the clinician only to the extent that these results are reported in an equally rapid fashion. This paper discusses an approach where both manual and automated results are transmitted via a laboratory computer system as soon as they are generated in the laboratory. For the Vitek AutoMicrobic System, on-line interfaces have been developed; this system operates in a manner similar to instruments in clinical chemistry or hematology laboratories. Systems for on-line editing of bacterial identification and antimicrobial susceptibility data are also discussed. It is concluded that computer systems capable of accomplishing instantaneous transfer of traditional and rapid microbiologic results are available and such systems are operating in some clinical microbiology laboratories.

Anti-Bacterial Agents↗
PubMed2001

Effect of HbE and HbH on HbA1C level by ionic exchange HPLC comparing to immunoturbidimetry.

BACKGROUND: The hemoglobin (Hb)A1C level is widely used to monitor diabetes mellitus patients. The N-terminal amino acid valine of its beta chain is glycated. The assay of HbA1C is based on differences in the charge, chemical and structural properties of the protein. METHODS: There are fully automated instruments available in clinical chemistry laboratory to assay HbA1C level. The effect of hemoglobinopathies was studied between an ionic exchange high-pressure liquid chromatography (HPLC) (Bio-Rad Laboratories, USA) and immunoturbidimetry (BM/Hitachi 912 with Roche HBA1CII, Germany-Japan) assay. The influence of high level HbF relative to the HbA1C level by ionic exchange HPLC is known. The effect of HbE and HbH to the HbA1C level by ionic exchange HPLC comparing to immunoturbimetry was examined. The evaluation was performed on 34 normal controls (A2A), 17 beta thalassemia traits (A2 upward arrow A), 36 HbE heterozygotes (EA), 37 HbE homozygotes (EE), 36 beta thalassemia/HbE (EF/EFA), 11 EABart's diseases (EABart's), 34 Hb H diseases (A2/CSAH) and 13 cord blood samples (FA). CONCLUSIONS: Hemoglobinopathies can impact on the assay of HbA1C level such as HbE and HbH to ionic exchange HPLC. Although not studied as yet, this effect may influence the other methods such as affinity chromatography.

Chromatography, High Pressure Liquid↗
PubMed2006

High-throughput confocal microscopy for beta-arrestin-green fluorescent protein translocation G protein-coupled receptor assays using the Evotec Opera.

Ligand-activated G protein-coupled receptors (GPCRs) are known to regulate a myriad of homeostatic functions. Inappropriate signaling is associated with several pathophysiological states. GPCRs belong to a approximately 800 member superfamily of seven transmembrane-spanning receptor proteins that respond to a diversity of ligands. As such, they present themselves as potential points of therapeutic intervention. Furthermore, orphan GPCRs, which are GPCRs without a known cognate ligand, offer new opportunities as drug development targets. This chapter describes a systems-based biological approach, one that combines in silico bioinformatics, genomics, high-throughput screening, and high-content cell-based confocal microscopy strategies to (1) identify a relevant subset of protein family targets, (2) within the therapeutic area of energy metabolism/obesity, (3) and to identify small molecule leads as tractable combinatorial and medicinal chemistry starting points. Our choice of screening platform was the Transfluor beta-arrestin-green fluorescent protein translocation assay in which full-length human orphan GPCRs were stably expressed in a U-2 OS cell background. These cells lend themselves to high-speed confocal imaging techniques using the Evotec Technologies Opera automated microscope system. The basic assay system can be implemented in any laboratory using a fluorescent probe, a stably expressed GPCR of interest, automation-assisted plate and liquid-handling techniques, an optimized image analysis algorithm, and a high-speed confocal microscope with sophisticated data analysis tools.

Algorithms↗
PubMed1990

Evaluation of the Titertek-Enterobac-Automated System (TTE-AS) for identification of members of the family Enterobacteriaceae.

The Titertek-Enterobac-Automated System (TTE-AS, Flow Laboratories, Meckenheim, FRG) was evaluated for identification of members of the family of Enterobacteriaceae using a total of 816 bacterial strains obtained from stock cultures and clinical specimens. A comparison of the results of the TTE-AS kit with conventional methods was conducted. Identifications were classified as correct, not identified (two or more taxa possible, identification score less than 80%, supplementary tests for final identification were not performed), and incorrect. Correct identification results were further subdivided according to the level of identification by species or biotype: (a) greater than or equal to 98%, (b) 90% to 97%, (c) 80% to 89%. When compared with conventional identification results, the TTE-AS system correctly identified 95.3% (778 of 816 strains), 90.0% (734 strains) belonging to category (a), 3.4% (28 strains) belonging to category (b), and 2.0% (16 strains) belonging to category (c). For 18 (2.2%) of the remaining strains, the system generated the correct identification result, but with an identification level less than 80% (low discrimination results). 11 strains (1.3%) were misidentified and the system failed to generate identification results with 9 strains (1.1%). Reactions within TTE-AS were reproducible with an estimated probability of erroneous test results of 1.7%.

Enterobacteriaceae↗
PubMed2000

An IBM PC-based system for experimental haemodynamic studies.

An IBM PC-based real-time data acquisition, monitoring and analysis system for experimental haemodynamic studies was developed. Comprehensive haemodynamic signals, such as aortic and left ventricular pressures, aortic and coronary blood flows, two segmental lengths, two segmental thicknesses, electrocardiogram and airway pressure, were acquired and monitored to assess cardiac function. The system performs computer-aided analysis and derivations on a number of haemodynamic parameters and cardiac function indices. The system has been tested and validated extensively over a number of series of experimental haemodynamic studies to investigate the effects of anaesthetic agents, cardiovascular drugs, and changes in loading on normal and critically ischaemic myocardium of anaesthetised laboratory subjects. Without this specialised and automated system, the analysis of the data acquired from the haemodynamic studies would be too time-consuming and could not be fully performed.

Blood Pressure↗
PubMed2000

Optimal control of molecular motion: design, implementation, and inversion.

This paper reviews recent theoretical and experimental developments aimed at controlling molecular motion using tailored laser fields. Emphasis is given to seeking optimal designs for the laser controls and optimal implementation of the controls in the laboratory. Optimization on both counts provides a rigorous, flexible, and physically attractive means for obtaining the best possible control over molecular motion under any specified conditions. The theoretical design and laboratory implementation of control are best effected by a closed-loop process that draws on observations of the evolving molecular sample to steer it toward the desired target. Going beyond control, similar closed-loop laboratory learning concepts may lead to automated molecular monitors for inversion to systematically identify details of molecular Hamiltonians.

Chemistry, Physical↗