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An accurate method for direct dual-energy calibration and decomposition.

We propose the use of conic and cubic surface equations (surfaces of second and third order) to directly approximate the dual-energy equations (the integral equations for the dual-energy log-signal functions, i.e., the negative logarithms of the relative detector signals, considered as functions of the basis-material component thicknesses of the object) and especially their inverses. These types of surface equations require a minimum number of calibration points, and their solutions are smooth, monotonic functions with the correct linear asymptotic behavior. The accuracy of this method is investigated and compared to that of conventional polynomial approximations, both for simulated and real calibration data, taken from two split-detector systems. These systems provide a more stringent test of our method than comparable dual-kVp systems, due to the greater nonlinearity of their log-signal and inverse functions. For these systems, we show that direct approximation of the inverse dual-energy equations using the simple eight-term rational form of the conic surface equation provides an extremely fast decomposition algorithm, which is accurate, robust in the presence of noise, and which can be calibrated with as few as 9 calibration points, or robustly calibrated, with a built-in accuracy check, using only 16 calibration points. Also, we show that extreme accuracy of approximation (to within less than 10(-6) in log-signal and 1 micron in material thickness) is theoretically attainable using the eighteen-term form of the cubic surface equation, which has a closed-form analytic solution. Finally, we consider the effects of noise on calibration accuracy, and derive simple formulas which relate the true and apparent root-mean-square (rms) accuracies. These formulas then allow the comparison of the true rms calibration accuracies of various surface approximations, considered as functions of the total calibration heat loading of the x-ray tube.

Algorithms↗

The importance of "like to like" ISI calibrations with freeze dried plasmas. European Concerted Action on Anticoagulation.

AIM: To assess reliability of like to like and cross species calibrations using two types of certified freeze dried plasma calibrants--artificially depleted of vitamin K clotting factors, and from coumarin treated patients. METHODS: Six ECAA national control laboratories provided certified values for the freeze dried plasmas in terms of the human plain international reference preparation (IRP) (BCT/441) with the manual prothrombin time technique. Eight other ECAA national laboratories determined international sensitivity index (ISI) values in full fresh plasma same species and cross species WHO calibrations against a low ISI human IRP (BCT/441) of the ECAA low ISI human thromboplastin and high ISI ECAA rabbit thromboplastin. Parallel calibrations were performed using the certified values. RESULTS: Calibrations on fresh plasmas of the human ECAA reference thromboplastin (stated ISI = 0.95) gave ISI of 0.957 against the human IRP and 1.66 against the rabbit IRP. The ECAA rabbit (stated ISI = 1.67) gave an identical value on the fresh plasma calibration v the human IRP. With freeze dried depleted plasmas certified in terms of the human IRP, the ISI of the ECAA human was 1.01, but the ECAA rabbit (stated ISI = 1.67) gave a low ISI of 1.47. The freeze dried coumarin plasmas gave an ISI of 0.943 for the ECAA human but only 1.493 for the ECAA rabbit. CONCLUSIONS: Fresh plasmas give reliable ISI when calibrating thromboplastins in same species and cross species calibrations. Freeze dried plasmas certified in terms of a single IRP, whether artificially depleted or of coumarin plasma origin, cannot be used for calibration of dissimilar thromboplastins.

Anticoagulants↗

The influence of the calibration mode of a laser fluorescence device on caries detection.

The aim of the present study was to assess the influence of the calibration mode of a laser fluorescence measurement device on caries detection. While positioning the probe on an occlusal carious lesion or stained fissure using a clamp, the occlusal surface of each of 62 teeth was covered with a light-body silicone impression material to facilitate a reproducible probe position. Then, measurements of the occlusal lesions were performed after standard and individual calibration at four non-carious areas on the buccal surface of each tooth. Mean laser fluorescence values of the occlusal reading point following standard calibration were about 4-5 units higher compared to values measured after individual calibration. After 1 week, readings after both standard and individual calibration were not different from baseline values (p > 0.05). During individual calibration, a mean range of about 2 units could be observed when comparing the readings of the occlusal reading point with respect to the four different areas of each buccal surface. The present study indicates that laser fluorescence readings are dependent on the mode of calibration. Moreover, for individual calibration, the probe position has to be recorded carefully. It is suggested that the same mode of calibration should always be used, when employing laser fluorescence measurements for longitudinal caries monitoring.

Calibration↗

INR calibration of Owren-type prothrombin time based on the relationship between PT% and INR utilizing normal plasma samples.

Prothrombin time (PT) is clinically important and is used to monitor oral anticoagulant therapy. To obtain PT results in international normalized ratio (INR), the current standardization procedure is complex and involves reference reagents. The PT of diluted plasma samples can be determined with a combined thromboplastin (the Owren-type procedure), but not necessarily with a plain thromboplastin (the Quick-type procedure). Owren-type PT procedures can therefore, as an alternative to the INR calibration, be calibrated with diluted normal plasma to give PT results in percent of normal PT activity (PT%). The present study explored if a plasma-based calibration of an Owren-type PT procedure can be used to obtain results in INR. The approach was to establish a relationship between PT% and INR by multi-center analysis of 365 samples from healthy individuals and patients on warfarin treatment. INR values were obtained by manual Quick-type reference procedure and PT% values by various automated Owren-type procedures. A relationship INR = (1/PT% + 0.018)/0.028 was found. A calibration procedure, based on the relationship, was investigated. Calibrators were the median PT of 21 normal plasma at dilutions representing 100%, 50%, 25%, 12.5% and 6.25% of normal PT activity. These were assigned INR values of 1.00, 1.36, 2.07, 3.05 and 6.36. Calibration of various Owren-type assays was repeatedly performed by 5 expert laboratories during 3 consecutive years. The INR values of certain lyophilised or frozen control plasmas were determined. The frozen control plasmas had externally assigned INR values according to WHO guide-lines. Within the laboratory, CV was typically below 3%. No appreciable difference among the results of the different laboratories or the three assay occasions was found. Externally assigned and INR values were essentially identical to those found. These and other results indicated that the calibration procedure was reproducible, precise and accurate. Thus, an Owren-type PT assay can be calibrated with normal plasma samples to give results in INR and the investigated calibration procedure can be proposed for this purpose.

Automation↗

A portable volume/flow calibrating syringe.

Two samples of a manual 3 L calibrating syringe which displays an electronically calculated FEF25-75% were evaluated to determine its suitability as a flow calibrator were evaluated. Room air was discharged into a manual spirometer system known to be accurate. The calibrator-determined values correlated very closely with the spirometer values over an FEF25-75% range of 0.4 to 9 L/sec. The differences between calibrator and spirometer FEF25-75% values were small (mean +/- 1 percent, greatest 3.7 percent) and of little importance clinically. This portable, simple to operate calibrating syringe provides accurate FEF25-75% and volume values. By adding flow calibrating capability to a recommended standard volume calibrating syringe, it will facilitate the routine calibration of spirometers in the laboratory and in the field. Since it uses displaced air, it can be used at altitude and with flow measuring instruments without the correction required for calibrators using CO2 cartridges.

Calibration↗

Excimer laser calibration system.

BACKGROUND: Excimer laser photoablation for refractive and therapeutic keratectomies has been demonstrated to be feasible and practicable. However, corneal laser ablations are not without problems, including the delivery and maintenance of a homogeneous beam. We have developed an excimer laser calibration system capable of characterizing a laser ablation profile. METHODS: Beam homogeneity is determined by the analysis of a polymethylmethacrylate (PMMA)-based thin-film using video capture and image processing. The ablation profile is presented as a color-coded map. Interpolation of excimer calibration system analysis provides a three-dimensional representation of elevation profiles that correlates with two-dimensional scanning profilometry. Excimer calibration analysis was performed before treating a monkey undergoing phototherapeutic keratectomy and two human subjects undergoing myopic spherocylindrical photorefractive keratectomy. Excimer calibration analysis was performed before and after laser refurbishing. RESULTS: Laser ablation profiles in PMMA are resolved by the excimer calibration system to .006 microns/pulse. Correlations with ablative patterns in a monkey cornea were demonstrated with preoperative and postoperative keratometry using corneal topography, and two human subjects using video-keratography. Excimer calibration analysis predicted a central-steep-island ablative pattern with the VISX Twenty/Twenty laser, which was confirmed by corneal topography immediately postoperatively and at 1 week after reepithelialization in the monkey. Predicted central steep islands in the two human subjects were confirmed by video-keratography at 1 week and at 1 month. Subsequent technical refurbishing of the laser resulted in a beam with an overall increased ablation rate measured as microns/pulse with a donut ablation profile. A patient treated after repair of the laser electrodes demonstrated no central island. CONCLUSIONS: This excimer laser calibration system can precisely detect laser-beam ablation profiles. The calibration system correctly predicted central islands after excimer photoablation in a treated monkey cornea and in two treated human subjects. Detection of excimer-laser-beam ablation profiles may be useful for precise calibration of excimer lasers before human photorefractive and therapeutic surgery.

Animals↗

A new experimental determination of the dose calibrator setting for 188Re.

UNLABELLED: Accurate activity measurements of radionuclides using commercial dose calibrators requires that the correct dial setting (or calibration factor) be applied. The dose calibrator setting for the medical radionuclide 188Re (as 188ReO4-) has been determined experimentally using solution sources prepared and calibrated at the National Institute of Standards and Technology (NIST). METHODS: The specific activity of two sources (in units of MBq/g) in the standard 5-mL NIST ampoule and in a 5-mL SoloPak dose vial were calibrated using 4pibeta liquid scintillation counting with 3H-standard efficiency tracing and gamma-ray/bremmstrahlung counting in the NIST "4pi" gamma ionization chamber on gravimetrically related sources. RESULTS: The newly determined settings for the NIST Capintec CRC-12 dose calibrator are (631+/-4) x 10 and (621+/-3) x 10 for the respective ampoule and dose vial geometries with an expanded (at a presumed 95% confidence level) uncertainty of 0.4%-0.5% in the activity determination. The setting for the dose vial geometry was independently confirmed using a Capintec CRC-15R at Cedars-Sinai Medical Center using sources calibrated against a NIST standard. CONCLUSION: These new settings result in activity readings 28%-30% lower than those obtained using the previously recommended setting of 496 x 10. This discrepancy most likely results from underestimating the total radiation yield from 188Re decay when calculating the dose calibrator response. This study emphasizes the need for experimental determinations of dose calibrator settings in the geometry in which the measurements will be performed.

Calibration↗

Calibration and validation of multiple regression models for stormwater quality prediction: data partitioning, effect of dataset size and characteristics.

Two main issues regarding stormwater quality models have been investigated: i) the effect of calibration dataset size and characteristics on calibration and validation results; ii) the optimal split of available data into calibration and validation subsets. Data from 13 catchments have been used for three pollutants: BOD, COD and SS. Three multiple regression models were calibrated and validated. The use of different data sets and different models allows viewing general trends. It was found mainly that multiple regression models are case sensitive to calibration data. Few data used for calibration infers bad predictions despite good calibration results. It was also found that the random split of available data into halves for calibration and validation is not optimal. More data should be allocated to calibration. The proportion of data to be used for validation increases with the number of available data (N) and reaches about 35% for N around 55 measured events.

Calibration↗

Effect of the calibration method of a laser fluorescence device for detecting occlusal caries in primary molars.

PURPOSE: The purpose of this study was to verify the influence of individual calibration on sound tooth surfaces and the number of readings on in vitro performance of a laser fluorescence (LF) device to detect occlusal caries in primary teeth. METHODS: After standard calibration (on the porcelain reference object), 72 clinically suspect sites on occlusal surfaces of 54 exfoliated or extracted (for orthodontic purposes) primary molars were assessed using the DIAGNOdent. First, after individual calibration (on a sound surface of each tooth), 3 readings were performed for each site. Subsequently, another 3 readings were taken, but without individual calibration on a sound surface of the tooth. After these assessments, sites were also evaluated with only one reading after individual calibration. Histological validation was performed as the gold standard. LF values, sensitivity, specificity, accuracy (number of correct diagnosis in both sound and diseased teeth) were calculated and compared using the McNemar change test. The area under receiver operating characteristics (ROC) curves was also compared. RESULTS: Readings with standard calibration only (mean= 11.7 +/- 10.6 SD) were significantly higher statistically than assessment with 3 readings after individual calibration (10.7 +/- 10.7) and with 1 reading (10.2 +/- 8.3) after individual calibration. Nevertheless, sensitivity, specificity, accuracy, and area under ROC curve did not change significantly. CONCLUSION: Absence of individual calibration does not affect the laser fluorescence device's performance in detecting occlusal caries in primary teeth.

Calibration↗

A calibration protocol for serum-based secondary standards.

In this procedure, tentative calibration values are assigned to secondary standards by analysis of that material with the routinely used instrument calibrated by use of the former lot of material. Aliquots of serum from patients are then analyzed by the routine method and an appropriate calibration method in which primary standards are used. If the patient-sera results by both methods are the same, the assigned calibration value is assumed to be correct; if not, the calibration value of the secondary-standard material is adjusted to produce agreement in patient-sera results between the two methods. Thus, calibration of the routine method is linked via the calibration method to primary standards as the criterion of accuracy. This approach corrects for matrix interference of serum-based secondary standards. All that is expected for the secondary standard to obtain an accurate result for patient sera is to produce an analytical signal (irrespective of the origin of the signal) that is equivalent to the calibration value. Differences were significant between the secondary-standard calibration value we determined by this procedure and that assigned by the manufacturer for bilirubin, calcium, phosphorus, chloride, CO2, creatinine, glucose, sodium, uric acid, and commonly measured enzymes.

Autoanalysis↗

Mechanical and physiological calibration of four cycle ergometers.

Mechanical and physiological calibrations were performed on four research-grade cycle ergometers. Ten subjects rode each ergometer twice in a randomized testing order. The subjects pedaled at 60 rpm for 5 min at each of three power outputs, i.e., 49, 98, and 147 W. Heart rate, metabolic, and perceptual data were obtained each minute. Prior to and immediately following these test rides, mechanical calibrations were obtained in duplicate. From the mechanical calibrations, Ergometer A was approximately 10% below actual values at each power output. Ergometer B demonstrated a variable error, with the largest percentage and absolute errors occurring at the lower power outputs. Ergometers C and D generally demonstrated less than a +/- 3% error. Following the physiological calibration, Ergometer B exhibited a substantial drift in calibration, while Ergometers A, C, and D maintained their original calibration. The physiological data supported the mechanical calibration, and Ergometer B demonstrated a substantial drift between the first and second trials and produced substantially different results compared with the other three ergometers. Ergometers A, C, and D demonstrated acceptable consistency in results both within trials and among ergometers. These results demonstrate the importance of proper calibration and of understanding the calibration characteristics of ergometers selected for research purposes.

Adult↗

Quantitative coronary arteriography: efficient correction of catheter calibrated vessel measurement.

1. BACKGROUND. To quantify coronary dimensions from digitized angiograms, the coronary catheter is commonly used as a scaling device. However, significant errors may result due to different angiographic magnification (DM) of the vessel and of the catheter resulting from deviating locations in the X-ray field. These errors can be corrected by biplane angiography. Since this correction needs a gantry measurement system, DM correction is less widely used. We analyzed the magnitude of DM that affects the accuracy of catheter calibrated vessel dimensions and developed DM corrections requiring low computational and measurement effort. 2. MATERIAL AND METHODS. We filmed biplane a perspex bloc of vessel phantoms (0.3 - 4.5 mm) in the isocenter of the x-ray system to get the true pixel sizes before any calibration. The phantoms were detected in the digitized images using a cardiac workstation (Kontron Cardio 500) and calibrated with differently located catheters. The calibration error was then calculated for increasing distances phantom-catheter. For DM correction, we developed procedures decreasing: (i) the computational effort (E) by approximated (A) formulas, as well as (ii) the measurement effort; this decrease approximates measured gantry settings (M) through fixed pre-settings for distances (D) and videochain (V). The several DM corrections were applied on the phantom images to analyze their correction performance and remaining calibration error. 3. RESULTS. The calibration of correctly detected pixel dimensions by a differentially magnified catheter causes a deviation of the absolute vessel dimensions, increasing with the vessel size. However, as shown in Table 1, DM correction reduces this calibration error substantially. Table 1: Calibration errors of catheter calibration and performance of DM corrections with decreasing effort (50 mm location distance phantom-catheter) 4. CONCLUSIONS. DM causes errors of vessel diameters of up to 15%. Even a measurement-free procedure (fixed presettings) corrects 71% of DM. Thus, the only additional effort of selecting an angiogram from the opposite view and marking the vessel and catheter locations in both images should be accepted.

Calibration↗

Application of a calibration method provides more realistic results for a finite element model of a lumbar spinal segment.

BACKGROUND: An important step in finite element modeling is the process of validation to derive clinical relevant data. It can be assumed that defect states of a finite element model, which have not been validated before, may predict wrong results. The purpose of this study was to show the differences in accuracy between a calibrated and a non-calibrated finite element model of a lumbar spinal segment for different clinical defects. METHODS: For this study, two geometrically identical finite element models were used. An in vitro experiment was designed, deriving data for the calibration. Frequently used material properties were obtained from the literature and transferred into the non-calibrated model. Both models were validated on three clinical defects: bilateral hemifacetectomy, nucleotomy and interspinous defects, whereas in vitro range of motion data served as control points. Predictability and accuracy of the calibrated and non-calibrated finite element model were evaluated and compared. FINDINGS: Both finite element models could mimic the intact situation with a good agreement. In the defects, the calibrated model predicted motion behavior with excellent agreement, whereas the non-calibrated model diverged greatly. INTERPRETATION: Investigating the biomechanical performance of implants and load distribution of different spinal structures by numerical analysis requires not only good agreement with the intact segment, but also with the defect states, which are initiated prior to implant insertion. Because of more realistic results the calibration method may be recommended, however, it is more time consuming.

Finite Element Analysis↗

The calibration and use of plane-parallel ionization chambers for dosimetry of electron beams: an extension of the 1983 AAPM protocol report of AAPM Radiation Therapy Committee Task Group No. 39.

This report is an extension of the 1983 AAPM protocol, popularly known as the TG-21 Protocol. It deals with the calibration of plane-parallel ionization chambers and their use in calibrating therapy electron beams. A hierarchy of methods is presented. The first is to calibrate the plane-parallel chamber in a high energy electron beam against a cylindrical chamber which has an Ncylgas value that has been obtained from a NIST traceable 60Co beam calibration. The second method, which is recommended for implementation by the ADCLs is an in-air calibration against a NIST-traceable calibrated cylindrical chamber in a Cobalt-60 beam to obtain a plane-parallel-chamber calibration factor in terms of exposure or air kerma. The third method places the two chambers in a phantom in a Cobalt-60 beam, and leads to an Nppgas value for the plane-parallel chamber. This report also gives Nppgas/NxAion)pp and Nppgas/(NkAion)pp values for five commonly used commercially available plane-parallel chambers: the Capintec PS-033, the Exradin P-11, the Holt, the NACP and the PTW-Markus. The calculation of these Ngas ratios introduces a Kcomp factor which is also calculated for the five parallel plate chambers. The use of the plane-parallel chambers follows the 1983 AAPM protocol for absorbed dose calibrations of electrons, except that new energy-dependent Prepl values are given for the Capintec PS-033 and PTW-Markus chambers consistent with the consensus of reports in the literature. For all the chambers, however, Prepl is unity for 20 MeV electrons. This report does not address the issue of the use of plane-parallel chambers in calibrating photon beams.

Cobalt Radioisotopes↗

The original calibre of the lower limbs arteries as a possible risk factor for complications of atherosclerosis: a statistical investigation in 90 subjects by echocolor-doppler.

Ninety subjects with emodynamically significant atherosclerotic disease of the lower limbs were examined. They had no history of diabetes mellitus or hypertension. Each subject underwent a color Doppler ultrasonographic study of the common iliac, superficial femoral, and popliteal arteries. In each arterial segment, diameter and blood flow velocity were measured. In evaluating the hemodynamic significance of the stenoses, we used the Windsor method. In a comparison of the calibers of the arteries significant relationship emerged in each given subject in two sexes. Males: a statistically significant difference was found only in the iliac artery, in which the calibre and Windsor indices were greater in the right as compared to the left; right: 1) There was a statistically significant relationship between mean caliber and Windsor indices, (p < 0.01); 2) there was a statistically significant correlation between mean calibre and age (p < 0.05); Left: 1) There was a statistically significant correlation between mean calibre and Windsor indices calculated at the levels of both the posterior tibialis (p < 0.05) and dorsalis pedis arteries (p < 0.01); 2) there was a statistically significant correlation between mean calibre and age (p < 0.05). Females: Student's t test for paired samples of the three arteries did not reveal a statistically significant predominance of one side over the other. With respect to the coefficients of correlation between mean calibre and Windsor indices, the results were as follows. Right: 1) There was a statistically significant correlation between mean calibre and Windsor indices calculated at the levels of both the posterior tibialis and dorsalis pedis arteries (p < 0.01); 2) there was a statistically significant correlation between age and mean calibre (p < 0.05); Left: 1) There was a statistically significant correlation between mean calibre and Windsor indices calculated at the levels of both the posterior tibialis (p < 0.05) and dorsalis pedis arteries (p < 0.01); 2) there was a statistically significant correlation between mean calibre and age (p < 0.05).

Aged↗

Achieving accuracy for routine clinical chemistry methods by using patient specimen correlations to assign calibrator values. A means of managing matrix effects.

Accurate results obtained from routine methods used in the clinical laboratory can be achieved if the methods are traceable to definitive or reference methods or are based on widely accepted methodologic principles. At Eastman Kodak Co (Rochester, NY), we assign calibrator values for each method that is available on Kodak Ektachem analyzers by correlation to a reference method or a methodologic principle using patient specimens. Kodak's Reference Laboratory is responsible for maintaining the accuracy and consistency of the reference method or methodologic principle by standardizing with National Institute of Standards and Technology materials, where available, monitoring performance through statistical process control, participating in proficiency testing programs, and following procedures consistent with the International Organization for Standardization guideline, "General Requirements for the Competence of Calibrations and Testing Laboratories (ISO 25). "Patient samples that span the reportable range are run in replicate on the reference method or methodologic principle over several days. These patient sample results are then used to create a calibration curve based on the relationship of concentration to the raw response of the routine method, where the same patient samples are also run in replicate on the same day. Each day, multiple vials of stable calibrator fluids are included in these tests, along with control materials. Each calibrator is assigned a value of equivalent concentration (in millimoles per liter) by predicting the concentration that would be present at the measured response if the test fluid were a patient sample. These are termed supplementary assigned values (SAVs). The same calibrators can be used to compensate for matrix effects resulting from changes in reagent formulations, by appropriately adjusting the assigned values. For example, when a process improvement for Kodak Ektachem clinical chemistry slides for phosphorus was introduced, an SAV change of 0.39 mmol/L (1.2 mg/dL) was required for one calibrator to maintain accuracy of the patient samples. A comparison of patient samples, using the SAVs specific for each formulation, gave excellent correlation (in millimoles per liter; r2 = 0.995) by the following equation: New Formulation = 1.02 (Old Formulation) -0.016; Sy.x = 0.074. The benefits of this approach to calibration include (1) accuracy is traceable to a reference method or methodologic principle and, wherever available, to reference materials; (2) new calibrator values (SAVs) can be assigned whenever the reagent formulation changes to maintain accurate patient results; and (3) if matrix effects are present, they will not adversely influence accuracy because traceability is based on patient sample comparisons.

Bias↗

RR-interval-based atrial fibrillation detection and burden estimation: cross-dataset validation and calibration-aware probability analysis.

Objective.Atrial fibrillation (AF) burden has become an increasingly important endpoint in long-duration rhythm monitoring, but reliable burden estimation requires more than accurate AF detection alone. In particular, when burden is derived by aggregating predicted AF probabilities over time, probability calibration may directly affect burden validity under external dataset shift.Approach.This study developed an interpretable-interval feature model for AF detection and evaluated it using record-wise cross-validation on a development cohort and independent cross-dataset external validation on public Holter electrocardiographic databases. Window-level performance was assessed using the area under the receiver operating characteristic curve (ROC-AUC), area under the precision-recall curve (PR-AUC), Brier score, expected calibration error (ECE), and calibration intercept and calibration slope. Recording-level AF burden was estimated using both probability-based and hard-label aggregation and evaluated using mean absolute error (MAE) and agreement analyses.Main results.The model showed high discrimination in both development and external evaluation, with external ROC-AUC ofand PR-AUC of. However, external calibration deteriorated despite preserved ranking performance, with Brier score of, ECE(15) of, calibration intercept of, and calibration slope of. In the external cohort, probability-based burden estimation preserved strong association with reference burden but showed weaker raw agreement than hard-label aggregation, with MAE ofversus, consistent with systematic probability underprediction. Repeated external recalibration across record-level splits substantially improved probability quality and probability-based burden estimation. Median probability-burden MAE decreased fromwithout recalibration toafter Platt recalibration andafter isotonic recalibration, while median ECE(15) decreased fromtoand, respectively.Significance.These findings indicate that-interval-based AF detection maintained strong ranking performance in the tested external cohort, but probability calibration should be evaluated explicitly when predicted probabilities are aggregated into AF-burden estimates.

Atrial Fibrillation↗

Use of flow cytometry and SNARF to calibrate and measure intracellular pH in NS0 cells.

BACKGROUND: Two calibration methods have been proposed for determining the relation between the fluorescence ratio of a pH-sensitive fluorescent indicator and intracellular pH (pHi). The first method uses nigericin to clamp pHi to external pH (pHe) and the second is the null point method. We compared these different calibration methods, solution conditions, and temperatures by using flow cytometry and the fluorescent dye 1,5- (and-6)-carboxy seminaphtorhodafluor-1-acetoxymethyl ester with an NS0 cell line. METHODS: The nigericin method was performed in glucose solutions supplemented with KCl and 2-(N-morpholino)ethane sulphonic acid plus tris(hydroxymethyl)aminomethane (solution 1A), a mixture of K2HPO4/KH2PO4 in glucose-solution supplemented solutions (solution 2A), or bicarbonate buffered growth medium supplemented with K2HPO4/KH2PO4 (solution 2B); this allowed a range of pHe values to be used. The effect of temperature (22 degrees C or 37 degrees C) on the nigericin calibration curve was also investigated. The null point method was performed by using a series of solutions with a mixture of weak acid and base with a known pHi response. RESULTS: Using solution 1A as the calibration solution resulted in acidic values of pHi for cells cultured in medium as compared with the values achieved with solution 2A. Using solution 2B did not affect the calibration curve. For the temperatures considered in this study, there was no affect on the calibration curve, but temperature did affect the pHi value of cells in phosphate buffered saline. The pseudo-null point method used with flow cytometry resulted in a calibration curve that was significantly different (P<0.05) from that achieved using the nigericin method. CONCLUSIONS: Our data indicates that the choice of calibration solution can affect the reported pHi value; therefore, careful choice of solution is important.

Animals↗