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PubMed2006

Automated high-throughput process for site-directed mutagenesis, production, purification, and kinetic characterization of enzymes.

Site-directed mutagenesis followed by functional characterization is a widely used approach to obtain information on the structure-function relationship of proteins. Due to time and cost considerations, the number of amino acids studied is frequently reduced. To address the need for convenient parallel production of numerous point mutants of a protein, we developed an automated method to perform classical site-directed mutagenesis, protein purification, and characterization in a high-throughput manner. The process consists of a succession of six fully automated protocols that can be adapted to any automated liquid handling systems. Our procedure allows construction, validation, and characterization of hundreds of site-directed mutants of a given protein in just 4 days. The method is especially adapted to projects aiming at the study of unique or multiple mutants without the need to construct and screen large libraries of random mutants. The usefulness of the technique is illustrated by the construction and characterization of tens of single mutants of the penicillin-binding protein 2x (PBP2x) from Streptococcus pneumoniae. Moreover, seven mutations of PBP2x were obtained simultaneously in a single experiment with efficiency close to 90%.

Enzymes↗
PubMed2006

Automated vs manual triage for bioterrorist disaster: a blinded crossover feasibility study comparing personal digital assistant to paper-based triage.

INTRODUCTION: This article reports results of a National Institutes of Health/National Library of Medicine Small Business Innovation Research-funded research grant comparing paper-based and automated Palm handheld computer disaster triage documentation. OBJECTIVES: The aim of this study was to test the feasibility of automated handheld computer triage and compare it to handwritten triage. METHODS: A paired t test was used in an intraindividual, blinded, crossover study to compare the 2 methods of disaster triage by 2 objective measures--time and accuracy. A total of 57 experienced, licensed first responders participated. Results are from analysis of 8 disaster scenarios with a total of 400 patients triaged using the 2 methods of documentation, crossed over, blinded, and paired per participant. RESULTS: The study demonstrated the feasibility of using TriageDoc, a Palm personal digital assistant (PDA ) program, as a viable alternative to current manual disaster triage. Furthermore, the PDA program gave advantage to bioterrorist agent identification. CONCLUSIONS: The feasibility of an automated Palm (Palm, Inc., Sunnyvale, CA) PDA triage program was demonstrated in this study. Study limitations, by the number of participants and the fact it is feasibility research, are acknowledged. Nevertheless, the research demonstrated TriageDoc was as accurate or more accurate as the manual method of triage with a tendency to require less time. Also there was no statistically significant difference between research sites with respect to accuracy or time to completion when the TriageDoc system was used. The program provided consistency and had flexibility in adapting to the various differences in triage methods at different locations. Hence, PDA programs such as TriageDoc may have potential advantages over handwritten documentation for disaster triage.

Bioterrorism↗
PubMed2006

Detection of epidermal growth factor receptor (EGFR) mutations in non-small cell lung cancer (NSCLC) using a fully automated system with a nano-scale engineered biomagnetite.

A fully automated system using nano-scale engineered biomagnetite was developed to detect mutations in the epidermal growth factor receptor (EGFR) gene in non-small cell lung cancer (NSCLC). Bacterial magnetic particles (BacMPs) were isolated from the magnetic bacterium Magnetospirillum magneticum AMB-1 and conjugated to streptavidin. Biotin-labeled target PCR products were then captured with the BacMPs, hybridized with the detection probe and detected by fluorescence signaling. The process was performed using a newly designed automated processor equipped with an XYZ mobile arm containing a 96-way automated pipetter, reagent dispenser and fluorescence detector. Two types of somatic mutations (in-frame deletions and point substitutions) in the EGFR gene were successfully identified within 3.5h using this system, suggesting that this system could be used in clinical tests of EGFR gene mutations in lung cancer, and potentially other cancer, patients. Additionally, a very low mutation rate could be detected in these samples.

Carcinoma, Non-Small-Cell Lung↗
PubMed2006

Evaluation of two fully automated novel enzyme-linked immunosorbent assays for the determination of human adiponectin in serum.

BACKGROUND: In contrast to RIA, recently available ELISAs provide the potential for fully automated analysis of adiponectin. To date, studies reporting on the diagnostic characteristics of ELISAs and investigating on the relationship between ELISA- and RIA-based methods are rare. METHODS: Thus, we established and evaluated a fully automated platform (BEP 2000; Dade-Behring, Switzerland) for determination of adiponectin levels in serum by two different ELISA methods (competitive human adiponectin ELISA; high sensitivity human adiponectin sandwich ELISA; both Biovendor, Czech Republic). Further, as a reference method, we also employed a human adiponectin RIA (Linco Research, USA). Samples from 150 patients routinely presenting to our cardiology unit were tested. RESULTS: ELISA measurements could be accomplished in less than 3 h, measurement of RIA had a duration of 24 h. The ELISAs were evaluated for precision, analytical sensitivity and specificity, linearity on dilution and spiking recovery. In the investigated patients, type 2 diabetes, higher age and male gender were significantly associated with lower serum adiponectin concentrations. Correlations between the ELISA methods and the RIA were strong (competitive ELISA, r=0.82; sandwich ELISA, r=0.92; both p<0.001). However, Deming regression and Bland-Altman analysis indicated lack of agreement of the 3 methods preventing direct comparison of results. The equations of the regression lines are: Competitive ELISA=1.48 x RIA-0.88; High sensitivity sandwich ELISA=0.77 x RIA+1.01. CONCLUSIONS: Fully automated measurement of adiponectin by ELISA is feasible and substantially more rapid than RIA. The investigated ELISA test systems seem to exhibit analytical characteristics allowing for clinical application. In addition, there is a strong correlation between the ELISA methods and RIA. These findings might promote a more widespread use of adiponectin measurements in clinical research.

Adiponectin↗
PubMed2006

Comparison of three automated urinalysis systems--Bayer Clinitek Atlas, Roche Urisys 2400 and Arkray Aution Max for testing urine chemistry and detection of bacteriuria.

BACKGROUND: Our study is aimed to determine the performance of 3 automated urinalysis systems-Clinitek Atlas, Urisys 2400 and Aution Max. METHODS: One thousand urine specimens were analyzed with the 3 automated systems. The results of the 3 assays were compared for testing urine chemistry and evaluating the capacity of leukocyte esterase and nitrite to detect bacteriuria. RESULTS: The correlation between the 3 instruments represented as within 1 grading difference was better between the Atlas and Aution Max systems for pH, blood, glucose, urobilinogen, ketone and specific gravity. For protein and nitrite, better correlation was observed between the Atlas and Urisys 2400, while the Aution Max and Urisys 2400 conveyed better correlation for bilirubin and white blood cells. The sensitivity and specificity of both the leukocyte esterase and nitrite in screening for significant bacteriuria were 71.7, 58.9, 70.8% and 99.1, 99.1 and 97.2%, for the Clinitek Atlas, Aution Max and Urisys 2400, respectively. CONCLUSIONS: The automated urinalysis systems demonstrate acceptable correlations with each other in urine chemistries, especially between the Clinitek Atlas and Aution Max systems on the majority of items. The specificity and negative predictive value of leukocyte esterase and nitrite of the 3 instruments for screening of significant bacteriuria were sufficient to avoid unnecessary urine culture.

Bacteriuria↗
PubMed2006

Automation and optimization of liquid-phase microextraction by gas chromatography.

Several fully automated liquid-phase microextraction (LPME) techniques, including static headspace LPME (HS-LPME) (a drop of solvent is suspended at the tip of a microsyringe needle and exposed to the headspace of the sample solution), exposed dynamic HS-LPME (the solvent is exposed in the headspace of sample vial for different time, and then withdrawn into the barrel of the syringe. This procedure is repeated a number of times), unexposed dynamic HS-LPME (the solvent is moved inside the needle and the barrel of a syringe, and the gaseous sample is withdrawn into the barrel and then ejected), static direct-immersed LPME (DI-LPME) (a drop of solvent is suspended at the tip of a microsyringe needle and directly immersed into the sample solution), dynamic DI-LPME (the solvent is moved inside the needle and the barrel of a syringe, and the sample solution is withdrawn and ejected), and two phase hollow fiber-protected LPME (HF-LPME) (a hollow fiber is used to stabilize and protect the solvent), auto-performed with a commercial CTC CombiPal autosampler, are described in this paper. Critical experimental factors, including temperature, choice of extraction solvent, solvent volume, plunger movement rate, and extraction time were investigated. Among the three HS-LPME techniques that were evaluated, the exposed dynamic HS-LPME technique provided the best performance, compared to the unexposed dynamic HS-LPME and static HS-LPME approaches. For DI-LPME, the dynamic process can enhance the extraction efficiency and the achieved method precision is comparable with the static DI-LPME technique. The precision of the fully automated HF-LPME is quite acceptable (RSD values below 6.8%), and the concentration enrichment factors are better than the DI-LPME approaches. The fully automated LPME techniques are more accurate and more convenient, and the reproducibility achieved eliminates the need for an internal standard to improve the method precision.

Chemistry Techniques, Analytical↗
PubMed2005

Determination of opiates and cocaine in hair using automated enzyme immunoassay screening methodologies followed by gas chromatographic-mass spectrometric (GC-MS) confirmation.

The objective of this study was to develop a two-step strategy for analysis of opiates and cocaine in hair samples involving an immunological screening procedure followed by confirmation of results using gas chromatography-mass spectrometry (GC-MS). A semi-quantitative automated competitive enzyme-linked immunosorbent assay (ELISA) methodology using Oral Fluid Micro-Plate Enzyme Immunoassays (Orasure Technologies, Inc.) was developed and validated. Applicability was proven by analysis of authentic head hair samples from drug users (n=103) and from opiate associated fatalities (n=21). The optimum cutoff values for the ELISA tests were 0.1 ng cocaine-equivalents/mg hair and 0.05 ng morphine-equivalents/mg hair using a 50 mg hair sample. Both ELISA tests had a sensitivity of 100%, the specificity was 66% for cocaine-equivalents and 42% for morphine-equivalents. The intraassay precision was 11% for the cocaine and 3% for the opiates ELISA, while interassay precision was 12% for the cocaine and 4% for the opiates ELISA test. The actual analyte concentrations in the hair samples were determined using GC-MS and were between 0.04 and 5.20 ng/mg for heroin (HER), between 0.04 and 30.01 ng/mg for 6-monoacetylmorphine (MAM), between 0.03 and 11.87 ng/mg for morphine (MOR), between 0.02 and 1.84 ng/mg for codeine (COD), between 0.02 and 2.48 ng/mg for acetylcodeine (AC), between 0.01 and 21.37 ng/mg for cocaine (COC), between 0.03 and 10.51 ng/mg for benzoylecgonine (BE) and between 0.05 and 1.26 ng/mg for cocaethylene (CE). The automated ELISA tests were proven to be valid screening procedures for the detection of cocaine and opiates in hair as confirmed by GC-MS. Screening methods provide rapid and inexpensive automated pre-test procedures to detect drugs in hair or other matrices. For forensic purposes screening therefore represents an ideal complement to routinely applied GC-MS procedures.

Case-Control Studies↗
PubMed2005

Automated border detection on contrast enhanced echocardiographic images.

BACKGROUND: Accurate determination of left ventricular ejection fraction (LV EF) is of paramount importance in the evaluation of patients with cardiovascular disease. Quantitative techniques for the automated calculation of EF exist however, the robustness of these techniques is dependent on adequate endocardial border definition and therefore are difficult to use in patients with limited images. We sought to combine the endocardial border enhancing effects of contrast echocardiography with an automated border detection technique to provide quantitative and accurate determination of LV EF. METHODS: Thirty-nine consecutive patients referred to nuclear cardiology for EF determination underwent radionuclide angiography followed by echocardiographic imaging using prototype software that allowed automated border detection during contrast infusion. RESULTS: Adequate LV cavity opacification with contrast was possible in 38/39 patients. The mean radionuclide EF was 50+/-16% (range 19-73). There was no statistically significant difference between the mean nuclear EF and averaged echocardiographically determined EF (51+/-18%). The mean bias was 0.6 with limits of agreement that were +15 and -14. CONCLUSION: This study demonstrated that prototype software successfully tracked the contrast enhanced endocardial border allowing accurate calculation of LV EF.

Adult↗
PubMed2004

Automated model-based organ delineation for radiotherapy planning in prostatic region.

PURPOSE: Organ delineation is one of the most tedious and time-consuming parts of radiotherapy planning. It is usually performed by manual contouring in two-dimensional slices using simple drawing tools, and it may take several hours to delineate all structures of interest in a three-dimensional (3D) data set used for planning. In this paper, a 3D model-based approach to automated organ delineation is introduced that allows for a significant reduction of the time required for contouring. METHODS AND MATERIALS: The presented method is based on an adaptation of 3D deformable surface models to the boundaries of the anatomic structures of interest. The adaptation is based on a tradeoff between deformations of the model induced by its attraction to certain image features and the shape integrity of the model. To make the concept clinically feasible, interactive tools are introduced that allow quick correction in problematic areas in which the automated model adaptation may fail. A feasibility study with 40 clinical data sets was done for the male pelvic area, in which the risk organs (bladder, rectum, and femoral heads) were segmented by automatically adapting the corresponding organ models. RESULTS: In several cases of the validation study, minor user interaction was required. Nevertheless, a statistically significant reduction in the time required compared with manual organ contouring was achieved. The results of the validation study showed that the presented model-based approach is accurate (1.0-1.7 mm mean error) for the tested anatomic structures. CONCLUSION: A framework for organ delineation in radiotherapy planning is presented, including automated 3D model-based segmentation, as well as tools for interactive corrections. We demonstrated that the proposed approach is significantly more efficient than manual contouring in two-dimensional slices.

Algorithms↗
PubMed2006

Feasibility of fully automated detection of fiducial markers implanted into the prostate using electronic portal imaging: a comparison of methods.

PURPOSE: To investigate the feasibility of fully automated detection of fiducial markers implanted into the prostate using portal images acquired with an electronic portal imaging device. METHODS AND MATERIALS: We have made a direct comparison of 4 different methods (2 template matching-based methods, a method incorporating attenuation and constellation analyses and a cross correlation method) that have been published in the literature for the automatic detection of fiducial markers. The cross-correlation technique requires a-priory information from the portal images, therefore the technique is not fully automated for the first treatment fraction. Images of 7 patients implanted with gold fiducial markers (8 mm in length and 1 mm in diameter) were acquired before treatment (set-up images) and during treatment (movie images) using 1MU and 15MU per image respectively. Images included: 75 anterior (AP) and 69 lateral (LAT) set-up images and 51 AP and 83 LAT movie images. Using the different methods described in the literature, marker positions were automatically identified. RESULTS: The method based upon cross correlation techniques gave the highest percentage detection success rate of 99% (AP) and 83% (LAT) set-up (1MU) images. The methods gave detection success rates of less than 91% (AP) and 42% (LAT) set-up images. The amount of a-priory information used and how it affects the way the techniques are implemented, is discussed. CONCLUSIONS: Fully automated marker detection in set-up images for the first treatment fraction is unachievable using these methods and that using cross-correlation is the best technique for automatic detection on subsequent radiotherapy treatment fractions.

Feasibility Studies↗
PubMed2026

Intervention Without Borders - an Automated Self-Guided AI-Enhanced Psychoeducation Intervention for Dementia Caregivers: Parallel-Group Randomized Waitlist-Controlled Trial.

OBJECTIVE: To examine whether a fully automated, self-guided intervention (PDC30) could improve caregiver well-being over a 1-month waitlist control in an international sample. DESIGN: Randomized waitlist-controlled trial. SETTING: Web-based platform accessible globally. PARTICIPANTS: 441 individuals responded to study promotion on the internet, of whom 274 from 43 countries met the study criteria and were randomized. Eligible participants were adults providing &#x2265;10 care hours weekly to community-dwelling relatives with dementia, scoring &#x2265;5 on Patient Health Questionnaire-9 (PHQ-9), and without recent caregiver intervention. INTERVENTION: Available 24/7, PDC30 is a self-guided, automated intervention consisting of a Guidebook, an AI-powered counseling chatbot, and interactive applications for cognitive-behavioral techniques, relaxation, and caregiver-recipient bonding. MEASUREMENTS: At baseline and follow-ups at 1, 2, and 3 months, depression was assessed by PHQ-9. Secondary outcomes were measured with validated brief versions of anxiety, burden, and positive gains. RESULTS: Intent-to-treat analysis using mixed-effects regression showed treatment x time2 effects on all outcomes except anxiety. At 1-month follow-up, coinciding with exclusive access to PDC30, intervention caregivers showed significant improvements in depression (d = -0.37), burden (d = -0.34), and positive gains (d = 0.42). The differences mostly disappeared after control participants received the intervention, while improvements in both groups were sustained thereafter. Participants reported using the website several times weekly, were generally satisfied with it, and found the chatbot most helpful. CONCLUSIONS: The effects on depression and other outcomes were consistent with those observed for in-person programs, suggesting the viability of well-designed automated intervention. The study demonstrates the feasibility, acceptability, and potential global health impact of PDC30.

Humans↗
PubMed2005

Automation of cDNA microarray hybridization and washing yields improved data quality.

Microarray technology allows the analysis of whole-genome transcription within a single hybridization, and has become a standard research tool. It is extremely important to minimize variation in order to obtain high quality microarray data that can be compared among experiments and laboratories. The majority of facilities implement manual hybridization approaches for microarray studies. We developed an automated method for cDNA microarray hybridization that uses equivalent pre-hybridization, hybridization and washing conditions to the suggested manual protocol. The automated method significantly decreased variability across microarray slides compared to manual hybridization. Although normalized signal intensities for buffer-only spots across the chips were identical, significantly reduced variation and inter-quartile ranges were obtained using the automated workstation. This decreased variation led to improved correlation among technical replicates across slides in both the Cy3 and Cy5 channels.

Animals↗
PubMedDate not supplied

Adequacy of evolving national standardized terminologies for interdisciplinary coded concepts in an automated clinical pathway.

PURPOSE: The purpose of this analysis was to determine the adequacy of evolving national standardized terminologies with regard to coded data elements (concepts) in an automated clinical pathway designed to drive adherence with the American College of Cardiology (ACC)/American Heart Association (AHA) Guidelines for Evaluation and Management of Chronic Heart Failure. METHOD: Concepts were identified in a previously developed automated clinical pathway and associated tools. Once identified, concepts were categorized according to the conceptual domains identified by Campbell et al. (1997). A review of evolving national standardized terminologies and coding systems was initiated to determine if the identified concepts had corresponding representation in one of these coding systems. Available codes were then evaluated for adequacy with respect to national guideline adherence measures put forth by the Centers for Medicare/Medicaid Services (CMS) and Joint Commission on Accreditation of Healthcare Organizations (JCAHO). RESULTS: The concept domain model put forth by Campbell et al. (1997) worked well for organizing concepts and for providing a useful framework for data analysis. Using our method, 260 unique pathway concepts were identified, of which, 91.9% (239) are represented by one or more of the standardized coding systems. Logical Observation Identifiers Names and Codes (LOINC) and SNOMED CT alone represented 86.2% of the concepts. Seventy percent (70%) of the clinical pathway concepts are represented using the Health Insurance Portability and Accountability Act (HIPAA) mandated national terminologies alone. Less than 50% of CMS and JCAHO guideline adherence concepts were found to have representation in the HIPAA mandated terminologies. The addition of Logical Observation Identifier Names and Codes (LOINC) and SNOMED CT improved representation up to 86.4%, but did not include representation of all concepts necessary for complete electronic monitoring of guideline adherence. CONCLUSIONS: Evolving national standardized terminologies provided matching terms for the majority of the data elements in the automated clinical pathway. Standard clinical terminologies with granular terms such as LOINC and SNOMED CT are required to represent the depth and detail of certain procedures and guideline-based care. Gaps exist in Health Insurance Portability and Accountability Act (HIPAA) mandated terminologies for representing interdisciplinary concepts in national adherence measures.

Computational Biology↗
PubMed2004

Methods for automated concept mapping between medical databases.

The retrieval and exchange of information between medical databases is often impeded by the semantic heterogeneity of concepts contained within the databases. Manual identification of equivalent database elements consumes time and resources, and may often be the rate-limiting technological step in integrating disparate data sources. By employing semantic networks as an intermediary representation of the native databases, automated mapping algorithms can identify equivalent concepts in disparate databases. The algorithms take advantage of the conceptual "context" embodied within a semantic network to produce candidate concept mappings. The performance of automated concept mapping was evaluated by creating semantic network representations for two test laboratory databases. The mapping algorithms identified all equivalent concepts that were present in the databases, and did not leave any equivalent concepts unmapped. The utilization of conceptual context to perform automated concept mapping facilitates the identification of equivalent database concepts and may help decrease the work and costs associated with retrieval and integration of information from disparate databases.

Algorithms↗
PubMed2004

Single-cell variability in growing Saccharomyces cerevisiae cell populations measured with automated flow cytometry.

Cell cultures normally are heterogeneous due to factors such as the cell cycle, inhomogeneous cell microenvironments, and genetic differences. However, distributions of cell properties usually are not taken into account in the characterization of a culture when only population averaged values are measured. In this study, the cell size, green fluorescence protein (Gfp) content, and viability after automated staining with propidium iodide (PI) are monitored at the single-cell level in Saccharomyces cerevisiae cultures growing in a batch bioreactor using an automated flow injection flow cytometer system. To demonstrate the wealth of information that can be obtained with this system, three cultures containing three different plasmids are compared. The first plasmid is a centromeric plasmid expressing under the control of a TEF2 promoter the S65T mutant form of Gfp. The other two plasmids are 2 microm plasmids and express the FM2 mutant of Gfp under the control of either the TEF1 or the TEF2 promoter. The automated sampling, cell preparation, and analysis permitted frequent quantification of the culture characteristics. The time course of the data representing not only population average values but also their variability, provides a detailed and reproducible "fingerprint" of the culture dynamics. The data demonstrate that small changes in the genetic make up of the recombinant system can result in large changes in the culture Gfp production and viability. Thus, the developed instrumentation is valuable for rapidly testing promoter strength, plasmid stability, cell viability, and culture variability.

Bioreactors↗
PubMed2004

Automated variable selection methods for logistic regression produced unstable models for predicting acute myocardial infarction mortality.

OBJECTIVES: Automated variable selection methods are frequently used to determine the independent predictors of an outcome. The objective of this study was to determine the reproducibility of logistic regression models developed using automated variable selection methods. STUDY DESIGN AND SETTING: An initial set of 29 candidate variables were considered for predicting mortality after acute myocardial infarction (AMI). We drew 1,000 bootstrap samples from a dataset consisting of 4,911 patients admitted to hospital with an AMI. Using each bootstrap sample, logistic regression models predicting 30-day mortality were obtained using backward elimination, forward selection, and stepwise selection. The agreement between the different model selection methods and the agreement across the 1,000 bootstrap samples were compared. RESULTS: Using 1,000 bootstrap samples, backward elimination identified 940 unique models for predicting mortality. Similar results were obtained for forward and stepwise selection. Three variables were identified as independent predictors of mortality among all bootstrap samples. Over half the candidate prognostic variables were identified as independent predictors in less than half of the bootstrap samples. CONCLUSION: Automated variable selection methods result in models that are unstable and not reproducible. The variables selected as independent predictors are sensitive to random fluctuations in the data.

Aged↗
PubMed2005

Automated criteria-based selection and analysis of fluorescent synaptic puncta.

The use of fluorescent probes such as FM 1-43 or synapto-pHluorin to study the dynamic aspects of synaptic function has dramatically increased in recent years. The analysis of such experiments is both labor intensive and subject to potentially significant experimenter bias. For our analysis of fluorescently labeled synapses in cultured hippocampal neurons, we have developed an automated approach to punctum identification and classification. This automatic selection and processing of fluorescently labeled synaptic puncta not only reduces the chance of subjective bias and improves the quality and reproducibility of the analyses, but also greatly increases the number of release sites that can be rapidly analyzed from a given experiment, increasing the signal-to-noise ratio of the data. An important innovation to the automation of analysis is our method for objectively selecting puncta for analysis, particularly important for studying and comparing dynamic functional properties of a large population of individual synapses. The fluorescence change for each individual punctum is automatically scored according to several criteria, allowing objective assessment of the quality of each site. An entirely automated and thus unbiased analysis of fluorescence in the study of synaptic function is critical to providing a comprehensive understanding of the cellular and molecular underpinnings of neurotransmission and plasticity.

Animals↗
PubMed2006

A computer vision-based automated Figure-8 maze for working memory test in rodents.

The benchmark test for prefrontal cortex (PFC)-mediated working memory in rodents is a delayed alternation task utilizing variations of T-maze or Figure-8 maze, which requires the animals to make specific arm entry responses for reward. In this task, however, manual procedures involved in shaping target behavior, imposing delays between trials and delivering rewards can potentially influence the animal's performance on the maze. Here, we report an automated Figure-8 maze which does not necessitate experimenter-subject interaction during shaping, training or testing. This system incorporates a computer vision system for tracking, motorized gates to impose delays, and automated reward delivery. The maze is controlled by custom software that records the animal's location and activates the gates according to the animal's behavior and a control algorithm. The program performs calculations of task accuracy, tracks movement sequence through the maze, and provides other dependent variables (such as running speed, time spent in different maze locations, activity level during delay). Testing in rats indicates that the performance accuracy is inversely proportional to the delay interval, decreases with PFC lesions, and that animals anticipate timing during long delays. Thus, our automated Figure-8 maze is effective at assessing working memory and provides novel behavioral measures in rodents.

Algorithms↗