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PubMed2006

An infrastructure for high-throughput microscopy: instrumentation, informatics, and integration.

High-throughput, image-based cell assays are rapidly emerging as valuable tools for the pharmaceutical industry and academic laboratories for use in both drug discovery and basic cell biology research. Access to commercially available assay reagents and automated microscope systems has made it relatively straightforward for a laboratory to begin running assays and collecting image-based cell assay data, but doing so on a large scale can be more challenging. Challenges include process bottlenecks with sample preparation, image acquisition, and data analysis as well as day-to-day assay consistency, managing unprecedented quantities of image data, and fully extracting useful information from the primary assay data. This chapter considers many of the decisions needed to build a robust infrastructure that addresses these challenges. Infrastructure components described include integrated laboratory automation systems for sample preparation and imaging, as well as an informatics infrastructure for multilevel image and data analysis. Throughout the chapter we describe a variety of strategies that emphasize building processes that are scaleable, highly efficient, and rigorously quality controlled.

Animals↗
PubMed1989

Rheumatoid factors determined by fluorescence immunoassay: comparison with qualitative and quantitative methods.

Fluorescence immunoassay (FIA), a relatively new technique for measuring rheumatoid factors (RF), is automated, quantitative, and calibrated against the Centers for Disease Control reference material for RF. We studied the FIA method in relation to a panel of RF methods, both qualitative [latex (LA) and sheep cell agglutination (SSCA)], and quantitative [nephelometry and enzyme-linked immunoassay (ELISA)]. Regression analysis revealed a highly significant correlation between FIA and either LA (r = 0.90) or nephelometry (r = 0.87). The correlation between FIA and either SSCA (r = 0.62) or ELISA (r = 0.67) was less strong. FIA had the highest sensitivity (91%) of all these methods; the specificity was 86%. FIA provides an accurate, sensitive, and specific measure of RF, and is a good alternative for laboratories wanting to replace titer methods with automated laboratory analysis.

Enzyme-Linked Immunosorbent Assay↗
PubMed1993

Collaborative evaluation of the Radiometer Sensititre AP80 for identification of gram-negative bacilli.

A multicenter trial of the Sensititre AP80 panel read on the Sensititre AutoReader (Radiometer America, Westlake, Ohio) for the automated identification of gram-negative bacilli was conducted with 1,023 clinical isolates (879 members of the family Enterobacteriaceae plus 144 nonenteric organisms). Assignment of taxa was based on the computer-assisted interpretation of the results of a series of reactions with fluorogenic enzyme substrates after 5 h of incubation, with an incubation interval of approximately 18 h used when indicated. Accuracy was determined initially by comparison with the results obtained with the API 20E or Rapid NFT system (Analytab Products, Plainview, N.Y.). Isolates showing discrepancies were identified by using conventional biochemical profiles. Identifications were available after 5 h of incubation for 918 isolates (90%). Agreements with reference results for members of the family Enterobacteriaceae were 95.3 and 92.5% at the genus and species levels, respectively, and for the nonmembers of the family Enterobacteriaceae, the agreements with reference results were 95.1 and 84.7%, respectively. The Sensititre AP80 panel was found to be simple and convenient to use, allowed for the testing of three isolates per panel, required minimal supplementary testing for completion of identification, performed in a reproducible fashion, and demonstrated an accuracy of same-day identification comparable to that reported for other automated systems. The AP80 panel appears well suited for routine use in the clinical microbiology laboratory as an automated means of identifying both members of the family Enterobacteriaceae and nonenteric gram-negative bacilli.

Bacterial Typing Techniques↗
PubMed1988

Automation and work-simplification in the urinalysis laboratory. A pilgrim's progress.

The evolution of the modern clinical laboratory has produced a gap between medical/scientific competence on the one hand and management skills on the other. Physician-managers need both sets of competencies. Concepts of operations management and cost accounting shape criteria for strategic decisions in technology improvement programs. Automation and work-simplification are key strategies for improving cost performance in clinical laboratories.

Accounting↗
PubMed1982

Comparison of three methods for determination of crude protein in meat: collaborative study.

A study was designed to compare the Kjel-Foss automated macro-Kjeldahl method and a block digestion-steam distillation method. The official AOAC Kjeldahl method was used as a reference procedure. Six products with a crude protein range of 10-30% were analyzed by 23 laboratories. Five laboratories analyzed the samples by the official AOAC method, 8 laboratories used the automated Kjel-Foss method, and 11 laboratories used the block digestion with steam distillation method. Standard deviations for each product and each method for both repeatability and reproducibility are given. The block digestion-steam distillation method has been adopted official first action.

Animals↗
PubMed1998

Automation of polymerase chain reaction-based systems for detection of hepatitis C virus RNA.

Polymerase chain reaction-based molecular assays are gaining increasing importance in the diagnosis and monitoring of infectious diseases. Over the past several years, the development and application of these techniques has initiated a revolution in the diagnosis and monitoring of hepatitis C infection. Presently, molecular assays are exclusively done in especially dedicated laboratories. Advances in automation will bring these technologies into routine diagnostic laboratories and will make them widely used.

Automation↗
PubMed2005

Performance evaluation of the latest fully automated hematology analyzers in a large, commercial laboratory setting: a 4-way, side-by-side study.

Gamma-Dynacare is a Canadian-based community laboratory partnership formed in the mid-1990s through the merger of 3 prominent Ontario medical diagnostic laboratories. Laboratory Corporation of America acquired an interest in the GD partnership in mid 2002. We service more than 10,000 community-based Canadian clinicians, hospital partners, and private clients with an integrated customer-focused system that includes specimen collection, transportation, and results reporting services. With more than 1,700 highly qualified medical, technical, and support staff and a network of laboratories, Gamma Dynacare aims to be at the forefront of technological innovation to better service the clinician base and ultimately deliver better patient care. We were looking for a hematology analyzer that would allow. (1) standardization throughout Ontario in our 4 largest sites and (2) better performance to effectively handle aged samples and minimize slide review. To select the best, most productive hematology analyzer for our environment, it was decided to perform a side-by-side comparison of the top hematology analyzers from Abbott (Cell-Dyn 3500), Beckman Coulter (LH 750), Bayer (Advia 120), and Sysmex (XE 2100), utilizing the same samples. CBC, differential and reticulocyte parameters were all evaluated according to CLSI (formerly NCCLS) and established hematology analyzer evaluation guidelines. We assessed each analyzer for precision, linearity, carryover, stability, differential capabilities, slide review rates, and throughput (clean bench studies). Two hundred samples were assessed for differential and morphology flagging on each analyzer using the reference 400 cell manual differential for comparison. Throughput was assessed by analyzing 700 consecutive samples representative of our workload mix. Stability studies at 24 hours showed that the Beckman Coulter LH 750 was least affected by EDTA, effect with minimal changes in the mean corpuscular volume (MCV) and hematocrit. Both the Bayer Advia 120 and Sysmex XE 2100 showed an elevation of the MCV (up to 5 fL) and Hematocrit over the 24 hours. Analysis of the 200 randomly selected patient samples showed that, while the false-negative rates on each of the instruments were comparable, there were significant differences in the false-positive rates. This has important implications for slide review rates. For our specimen mix, the Sysmex XE 2100 had the highest false-positive rate (15%), followed by the Cell-Dyn 3500 (8%), Advia 120 (6.5%) and the Beckman Coulter LH 750 (1.5%). Reticulocyte analysis performance was observed to be satisfactory with the Beckman Coulter LH 750, Cell-Dyn 3500, and XE 2100, while the Bayer Advia 120 showed a decrease in retic values after 12 hours. In conclusion, many laboratories will not be able to perform a 4-way evaluation such as described here due to time, space, and resource constraints. For our laboratories, result quality, sample stability performance, slide review rates, and efficiency were the primary criteria in selection of the most suitable hematology analyzer. Our 4-way evaluation resulted in selection of the Beckman Coulter LH 750 for Gamma-Dynacare Laboratories because it enabled the lowest slide review rate and handled aged samples better than the other analyzers.

Automation↗
PubMed2005

Automated reporting of GFR--coming soon to a laboratory near you!

BACKGROUND: Serum creatinine concentration is an unreliable and insensitive marker of chronic kidney disease (CKD). To improve CKD detection, Australasian guidelines have recently recommended that laboratories calculate and report an estimated glomerular filtration rate (eGFR) using the Modification of Diet in Renal Disease (MDRD) formula with every request for serum creatinine concentration. OBJECTIVE: This articles aims to provide timely information to health professionals about how to appropriately interpret and act upon eGFR reports. It also discusses the treatments shown to reduce renal and cardiovascular risk in CKD patients, and the indications for nephrologist referral. DISCUSSION: The accuracy and precision of eGFRs are reasonable in most adults in whom calculated values are ENTITY lt 60 mL/min/1.73 m2. However, eGFRs should be interpreted with caution in some settings (particularly patients with eGFRs ENTITY gt 60 mL/min/1.73 m2 and children). Automatic laboratory reporting of eGFR will enhance early detection of CKD, allow the timely institution of appropriate reno- and cardio-protective therapies, and better inform decisions regarding the prescription of renally excreted medications.

Adult↗
PubMed1988

Breast cancer estrogen receptor assays: assessment of a direct hyperbolic fitting to analyse multipoint binding data.

Analysis of breast cancer estrogen receptor multipoint binding assay is performed by fitting experimental data to a hyperbolic model derived from the law of mass action. The calculations performed on a microcomputer are carried out from the total bound and free ligand concentrations. The parameters estimated by hyperbolic fitting, receptor concentration N and constant of dissociation K, well agree with those obtained by Scatchard's transformation. N and K derived from hyperbolic analysis are much less susceptible to the influence of experimental errors. The method is more reliable at low receptor concentrations. The main advantage of the hyperbolic fitting is to simplify the technical methodology in clinical laboratory practice; there is no need to determine the non-specific bindings experimentally. Calculations can be easily automated on any laboratory microcomputer. Assays of any kind of receptor could be analysed by the hyperbolic fitting when the physical-chemical equilibrium between receptor, nonsaturable component and ligand can be approximated by a two-component model.

Breast Neoplasms↗
PubMed1980

Characterizing blood cells by biophysical measurements in flow.

One effect of automation in the hematology laboratory has been to introduce new characterizations of blood cells. Resistive pulse sensing (Coulter) and light scatter measurements in flow provide rapid and reproducible cell counts. They also provide information about red cell size, shape, and deformability. Thus, they have provided new characterization of these cells in terms of their biophysical properties. Leukocytes have been classified by optical scatter and absorption measurements in flow after being stained cytochemically. This provides rapid and precise WBC differential counts. However, here again, additional information about relative cell-enzyme content or activity is also accessible to provide a new characterization of the leukocytes. The ultimate range of utility of this expanding technology in the automated hematology laboratory of the future will, of course, depend upon establishing relations between the biophysical parameters and the functions of the cells. This, in turn, must depend upon the use of the technology by researchers and clinicians in studying cell function and the aberrations of these functions which define disease.

Autoanalysis↗
PubMed2003

Evaluation of haematology analyzer CELL-DYN 3700 SL.

Research on the parameters of full blood count and differential white blood count is included in the program of all medical laboratories of primary, secondary and tertiary health care levels. Today, all haematological tests are exclusively performed on the haematology analyzers. Automation of haematology laboratories is a result of the huge requires for haematological test performing, timely issuing of the haematological findings, and possibility of the usage of modern techniques. This work is an evaluation of laser haematology analyzer Cell-Dyn 3700 SL. It investigates the reliability of test results throughout the following parameters: precision, accuracy, sensitivity and specificity of determination methods. It also explores the influence of sample transferring and correlation with haematology analyzer MAXM Retti. Haematology parameters that have been investigated are: white blood cell (WBC), neutrophils (NEU), lymphocytes (LXM), monocytes (MONO), eosinophils (EOS), basophils (BASO), red blood cells (RBC), haemoglobin (HGB), haematocrit (HCT), mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCHC) red cell distribution width (RDW), platelet (PLT), mean platelet volume (MPV), plateletocrit (PCT), and platelet distribution width (PDW). The results confirms that precision of analyzer fulfils the reproducibility of testing parameters: WBC, RBC, HGB, MCV, MCH, MCHC, and PLT. Correlation coefficient values (r) gained throughout the statistical analysis, that is linear regression results obtained throughout the comparison of two analyzers are adequate except for MCHC (r = 0.64), what is in accordance with literature data. Accuracy is tested by haematology analyzer method and microscopic differentiating method. Correlation coefficient results for granulocytes, lymphocytes and monocytes point the accuracy of methods. Sensitivity and specificity parameters fulfil the analytical criteria. It is confirmed that haematology analyzer Cell-Dyn 3700 SL is reliable for the determination of full blood count in everyday work. Analyzer and its program for differential white blood count can be used for the research and separation of normal and pathological blood counts with addition of microscopic methods confirming distribution or morphologic changes of leukocytes.

Journal Article↗
PubMed2004

The investigation of short stature: a survey of practice in Wales and suggested practical guidelines.

AIM: To survey the investigation of short stature in Wales and suggest guidelines to improve practice. METHODS: Questionnaires were circulated to paediatricians and consultant clinical biochemists or consultant chemical pathologists at 13 Welsh hospitals where children with short stature are investigated. RESULTS: A 100% response was obtained from laboratory and clinical staff. Clinicians screened 1-50 patients each year (median, 10). Growth hormone (GH) deficiency was subsequently diagnosed in 0-30% (median, 10%) and GH treatment started in 30-100% (median, 100%) of patients. Five paediatricians and eight laboratories had written investigative protocols. Investigation of GH secretion was initiated in some centres before a complete clinical evaluation was carried out. Various screening tests for GH deficiency, including insulin-like growth factor 1 (IGF-1), random GH, and exercise tests were used. The clonidine stimulation test was used to assess the GH axis in most centres but eight different protocols were described. GH was measured in four Welsh laboratories using two automated immunoassay methods. However, nine different ranges of cutoff values for defining abnormal GH responses were quoted, and in three centres laboratories and paediatricians quoted different cutoffs. CONCLUSION: This survey demonstrates the need for practical guidelines for the investigation and management of short stature in children, agreed by paediatricians and their laboratory colleagues. The guidelines should encompass the initial clinical investigation, assessment of the GH-IGF-1 axis (using standardised protocols), and provision for the transition to adult management. This article presents practical guidelines based on published points for good practice.

Child↗
PubMed1994

Mycobacterium abscessus pseudoinfection traced to an automated endoscope washer: utility of epidemiologic and laboratory investigation.

After 15 patients had positive cultures for Mycobacterium abscessus without evidence of infection (i.e., pseudoinfection) following endoscopy, retrospective cohort studies of patients undergoing endoscopy and microbiologic sampling of the environment were done to examine potential nosocomial transmission and to identify the source and risk factors for M. abscessus pseudoinfection. In the epidemic period, M. abscessus-positive cultures were significantly more likely to be obtained during bronchoscopy than gastroendoscopy (16/149 vs. 1/860, P < .001) and during procedures using bronchoscopes disinfected in an automated washer rather than by other methods (16/54 vs. 0/95, P < .001). M. abscessus was recovered from the automated washer, the inlet water feeding the washer, and a flexible bronchoscope. Environmental and case-patient isolates had identical large restriction fragment (LRF) patterns of genomic DNA separated by pulsed-field gel electrophoresis. Molecular typing using LRF analysis supported the epidemiologic results and demonstrate the utility of combined epidemiologic and laboratory investigations in nosocomial outbreaks of nontuberculous mycobacteria.

Adult↗
PubMed2004

The impact of automation on organizational changes in a community hospital clinical microbiology laboratory.

The diagnosis of infectious diseases and the role of the microbiology laboratory are currently undergoing a process of change. The need for overall efficiency in providing results is now given the same importance as accuracy. This means that laboratories must be able to produce quality results in less time with the capacity to interpret the results clinically. To improve the clinical impact of microbiology results, the new challenge facing the microbiologist has become one of process management instead of pure analysis. A proper project management process designed to improve workflow, reduce analytical time, and provide the same high quality results without losing valuable time treating the patient, has become essential. Our objective was to study the impact of introducing automation and computerization into the microbiology laboratory, and the reorganization of the laboratory workflow, i.e. scheduling personnel to work shifts covering both the entire day and the entire week. In our laboratory, the introduction of automation and computerization, as well as the reorganization of personnel, thus the workflow itself, has resulted in an improvement in response time and greater efficiency in diagnostic procedures.

Automation↗
PubMed1978

[Automation of chemical analysis in enology].

Automatic dosages took place a short time ago in oenology laboratories. First researchs about automation of usual manual analysis have been completed by I.N.R.A. Station of Dijon during 1969--1972 years. Then, other researchs were made and in 1974 the first automatic analyser appeared in application laboratories. In all cases continuous flow method was used. First dosages which has been carried out are volatic acidity, residual sugars, total SO2. The rate of work is 30 samples an hour. Then, an original way for free SO2 was suggested. At present, about a dozen of laboratories in France use these dosages. The ethanol dosage automation, very important in oenology, is very difficult to carry out. A new method using a thermometric analyzer is tested. Research about many dosages as tartaric, malic, lactic acids, glucose, fructose, glycérol, have been performed especially by I.N.R.A. Station in Narbonne. But these dosages are not current and at present no laboratory apply them. Now, equipments price and redemption, change of tradionnal dosages for automatical methods and the level of knowledge required for operators are well known. The reproducibility and the accuracy of the continuous flow automatic dosages allow, for enough important laboratories, to make an increasing number of analysis necessary for wine quality control.

Autoanalysis↗