Facscalibur manual

For those who prefer to work with Instrument setup is as easy as the push of a button and a click of the mouse.

Efficient collection optics, coupled to the flow cell with an optical gel, permit the use of a low-powered, air-cooled argon laser and a red diode laser to ensure the high sensitivity required for analysis of While performing any of a wide range of supported applications, the BD FACSCalibur system provides the tools to obtain results quickly, easily, and accurately. For over twenty-five years, BD has actively worked with researchers to develop tools that help improve workflow, ease of use, and performance.

For those who prefer to work with multiwell plates, BD Biosciences offers the first walkaway sample introduction device for and well plates. Sorting on the BD FACSCalibur occurs in a completely enclosed, aerosol-free environment for enhanced safety which is especially important when processing biohazardous samples. The sorting process is designed to be straightforward. After the population of interest is gated, simply click Acquire to begin the sort. After the sample is acquired and cells pass through the laser, a unique catcher tube mechanism moves in and out of the sample core stream at a rate of roughly times per second to capture designated cells and direct them to a collection tube or to an optional cell concentrator module for further processing.

While performing any of a wide range of supported applications, the BD FACSCalibur system provides the tools to obtain results quickly, easily, and accurately. The BD FACStation system automates many software functions and delivers high-performance acquisition and analysis tools for creating plots, gating, statistical analysis, and reporting. It performs all the computing tasks required for fast, accurate results, including instrument setup, data collection, analysis, and management.

For clinical applications, BD Biosciences offers a wide variety of cellular controls to verify performance for immunophenotyping, reticulocyte analysis, stem cell enumeration, and rWBC applications.

Signal Processing Workstation resolution:1, channels on all parameters. Fluorescence compensation network Width and area measurements for discriminating doublets; available for all fluorescence parameters.

Data entry: Sample information, reagent panels, and rack information can be defined for up to tubes 40 tubes x 16 racks at a time. Participating site staff routinely performed flow cytometric enumeration of hematopoietic stem cells and evaluated specimens for eligibility in this study. For frozen specimens, enrollment and staining were to be performed as soon as practicable and within one hour of thaw.

When required, WBC counts were adjusted by dilution just prior to staining in cold phosphate buffered saline PBS with 0. Normal peripheral blood NPB and mobilized peripheral blood MPB samples were required to be remnant material from routine laboratory testing and anticoagulated with ethylenediaminetetraacetate EDTA.

Only leftover specimens, defined as remnants of human specimens not used for a test, which would otherwise have been discarded, were evaluated for eligibility. On the day of the study, instrument setup was completed prior to running process controls.

Process controls achieved acceptable results prior to specimen enrollment. Specimens were prepared as duplicate samples 4 tubes for both investigational methods.

For the predicate, specimens were prepared as duplicate samples, along with one isoclonic control 3 tubes. NH 4 Cl solution 1X, 2 mL was added, and the tubes were vortexed and incubated for another 10 minutes in the dark. At the end of the lysing period, samples were transferred to melting ice, protected from light, and acquired within one hour.

Samples were re-vortexed immediately before acquiring. The tube was vortexed and incubated for 10 minutes in the dark. The target number of specimens was a minimum of enrolled specimens with valid results for each investigational method. Results were valid when the data were accurate, complete, and in compliance with all applicable protocol requirements. To be evaluated as part of the analysis sample pool and meet the primary endpoint for each investigational method, three prospectively determined criteria had to be satisfied.

To prevent inter-site variability due to inadequate technical proficiency, training of staff at each site was performed by BD Biosciences staff, and was followed by technical qualification and equivalency analysis between the sites.

Fresh LPH was used to show site equivalence on procedures for sample preparation, testing, and analysis. Determination of equivalency of each site was required to allow for poolabilty of the data to analyze the study dataset in its entirety, independent of the site at which each sample was analyzed. Prior to final analysis, data were reviewed, and some enrolled specimens were deemed not to be evaluable because they did not meet the eligibility requirements to allow entry into the dataset.

The most common reasons for exclusion from analysis were non-compliance with inclusion criteria or protocol procedures. Fresh samples were enrolled and stained within 24 hours post-draw. Frozen specimens were enrolled and stained as soon as practicable and within 1 hour post-thaw. If a specimen met all criteria, the enrollment process was documented on the appropriate case report form CRF and the specimen was delinked from the diagnostic specimen and or subject identification by assignment of a study-specific specimen number.

To further evaluate the robustness of the investigational as well as predicate methods, we compared the results for each site by specimen type and method. It is important to note the limitations of this analysis, since the specimens evaluated at each site were analyzed only at that particular site, and in no case was an analysis of a single sample performed at more than one site.

Therefore, direct comparability analysis between the sites was confounded by the difference in specimens at the site, since the specimens might be procured and processed following procedures affected by local practices. Therefore, of particular interest are the data generated from the analysis of specimen types that were less readily influenced by these local practices, such as NPB and fresh CB.

The concept of accuracy includes both bias systematic error and precision random error , where bias is defined as a persistent positive or negative deviation of the measured value from the true, or in this case predicate, value.

To evaluate agreement of the test method with the predicate, we devised an experimental design to evaluate the mean bias or difference terms that are used interchangeably in each of the cell-concentration bins, after control of random error through training of analytical personnel, test site qualification, demonstration of site equivalency, and required operational flow to balance various testing patterns.

Further, we balanced the number of sample types collected at each testing site involved in the study. Prior to analysis, site equivalency ie, poolability was evaluated using the mean bias of the LPH specimen from each site for each study method before pooling all data across sites.

For each investigational method, the mean bias between all sites must have fallen within the poolability criteria for site equivalency. All bias and regression analyses were performed using measured versus reported values of the investigational methods versus measured values of the predicate method.

The measured and reported values differ if specimens were diluted during sample preparation. The reported value refers to the measured value multiplied by the dilution factor. Concordance analyses were performed on reported values. Data from the first replicate of the investigational methods from each specimen were used for the bias calculation between the investigational and predicate methods.

Data from the second replicate of the investigational methods from each specimen were used for the outlier determination. These optimized results were used for endpoint analysis of this investigational method. The absolute difference and the relative difference were calculated using the following formulas:. Study testing was performed on a total of 1, remnant enrolled specimens that had been delinked of their patient identification through the application of new labeling to replace previous identifiers.

Of the total of 1, leftover and delinked specimens enrolled in the four clinical sites, the distribution of required and analyzed specimens by investigational method is shown in Table 2. The distribution of the required and analyzed specimens by investigational method, anticoagulant, and specimen type used in the study was well balanced between the two instruments. To be analyzed, matching evaluable results must have been available for the predicate method. Fresh specimens within 24 hours of collection and frozen specimens within 1 hour of thawing were enrolled in the study.

During data cleaning, specimens were eliminated from analysis because they did not meet the data evaluability criteria or because during testing the protocol procedures were not followed. The reasons for exclusion were failure to meet inclusion criteria, inappropriate instrument setup or sample preparation, poor review or lack of review of the data of quality control criteria after acquisition, and instrument errors.

Of the specimens excluded, 81 specimens were completely excluded from the analysis because the specimen was disqualified from the predicate method. Fresh LPH was used to show site equivalency on procedures for sample preparation, testing, and analysis. A minimum of 55 fresh LPH specimens were enrolled by site see Table 6. The bias analyses of Abs CD34 were grouped in low, mid, and high bins to obtain the absolute and relative differences from the predicate in both investigational methods.

Results are summarized in Table 7. Skip to main content Skip to navigation. You are now leaving the BD Biosciences website. The site you are about to visit is operated by a third party. The link to this site neither makes nor implies any representation or warranty for any products or services offered on a third-party site and is intended only to enable convenient access to the third-party site and for no other purpose. Do you want to continue?