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The output of the analog peak and hold circuitry is digitized using bit ADCs and placed on a patented parallel processing digital bus. The bus handles real-time compensation as well as other digitized signals, including area and width of up to eight additional pulses.
To sort cells, researchers can select subpopulations through the creation of up to 32 sort regions. A variety of sort modes, optimized to favor purity, yield, or count accuracy, is standard. A custom mode allows researchers to optimize those settings for a specific application. If purity is of no concern, theoretically sort speed is as fast as the throughput rate. In this case, cells are sorted independently and the target population is enriched.
In most sorts, however, purity is a major concern, and factors such as droplet formation rate, event rate, and sample quality play a deciding role. Accurately calculating the drop-delay determination—the point at which the drop separates from the stream—is important because it ensures that the instrument precisely places the charge on the drop containing the particle of interest. Accudrop technology assists the researcher to set the optimal drop-delay value.
The researcher can identify the precise drop-delay value without a fluorescence microscope or manual calculations. The BD Influx system can adapt to virtually any environment. A small footprint allows the instrument to fit into tight spaces, and a modular component design allows the instrument to be easily configured to specific needs. Footprint and modularity have made the BD Influx cell sorter the instrument of choice for marine biology applications.
The BD Influx Mariner version includes pre-amp modifications for dimly fluorescent microorganisms and a brace to secure fluidics tanks for shipboard installation. A range of standard detection modules is available to configure the system based on application needs. Optional, exchangeable detector modules provide the capability to measure small particles, the polarization state of scatter, or fluorescence signals.
This flexibility allows the user to adapt the system to the needs of routine or emerging applications. The BD Influx system can be equipped with polarization-sensitive detectors. A unique polarization design uses two detectors mounted under Brewster angles to measure changes in parallel and perpendicular light for both scatter or fluorescence. Changes in polarization can help differentiate between organisms containing highly reflective inclusions such as diatoms, or discriminate among different populations of granulocytes.
This option lowers the threshold on size measurements using a special detector with a high NA microscope lens, a pinhole, and a photomultiplier tube PMT.
The pinhole reduces the amount of stray light reaching the detector and prevents saturation of the PMT. Optimized sheath fluid 0. This resolution makes the Small Particle Option particularly valuable for applications in marine biology, microbiology, and environmental biology. The Small Particle Option also allows researchers to detect fluorescence in the forward direction. This capability is used in applications such as chlorophyll detection. For precise sorting, the mechanics and speed of the XY table in the Computerized Cell Deposition Unit CCDU allow sorting into microtiter plates or other custom devices such as slides or petri dishes.
Index sorting functionality has been completely rewritten and extended to put a very powerful analytical technique in the hands of researchers. It is now possible to review the complete flow phenotype of every cell sorted into a multiposition sort device, such as a well tray. Index sort mode creates an FCS file containing all the sort deposition information and tray position information on an event-by-event basis. Post-sorting results can be precisely traced back to the flow characteristics of the specific cell or combinations of cells sorted.
In addition to sorting into tubes, the BD Influx sorter supports an endless variety of slides, plates, and custom devices. Predefined trays and slides include , , , and 6-well plates, coarse and fine calibration slides, and 4 x 12 slides. Biological safety in flow cytometry is an emerging requirement for core laboratories concerned about the potential accidental exposure of operators to biological samples.
BSCs are designed to protect operators from risks associated with exposure to biological agents in samples.
These cabinets are among the most effective and commonly used primary containment devices in laboratories working with infectious agents. BSCs protect personnel and the environment from harmful agents and protect product cells from contamination. Importantly, all microbiological testing was performed with the BD Influx instrument placed inside the work area of the BSC to validate performance in an as-used condition.
The BSC controls the direction, volume, and speed of airflow to direct potentially harmful particles away from the operator. Air is filtered and circulated around the work surface, and a separate airflow at the front of the cabinet creates a protective barrier for the operator.
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The HEPA filters remove microorganisms and airborne particulates aerosols from the air. Filters are placed where the air enters and exits the work area. Filters are placed where a percentage of the air is exhausted from the cabinet and where the cabinet air is recirculated.
HEPA filters in the cabinet support the removal of a minimum of BD Influx. The software captures all the information about an event such as time, position in the sorted drop, position relative to other events, firmware classifier status, etc, and makes it available to researchers on demand as needed for quality control, or for post-sort analysis.
The software uses industry-standard protocols and manages hundreds of system parameters, giving researchers a higher level of control and engagement with the instrument to support advanced research applications.
To maximize reproducibility and accelerate study, researchers can save and recall previous workspaces, configurations, instrument states, and parameter settings. A multitasking capability allows researchers to sort cells from a sample, record the information in a data file, and work with a previous data file—all at the same time. Software wizards and controls assist researchers to classify cell populations, perform compensation, monitor sorting, and analyze results. Hierarchical gating tools make it easy and intuitive to classify cell populations.
The automatic compensation wizard creates a spillover matrix based on user-selected population controls and automatically calculates a compensation matrix to align populations. Sort controls and event counters monitor sorting, with the ability to pause, resume, reset, and stop the sort streams individually or all at once. Data and sort analysis tools provide robust statistics on cell populations and sorting quality control at the individual cell level.
Output formats include histograms, overlay histograms, and dot, density, and contour plots—all with linear, log, or biexponential Logicle scaling. Results can also be exported and imported as FCS data files for other software applications. Researchers can perform cytometer setup, compensation, data acquisition, gating, analysis, and sorting progressively— or they can choose to return to any step for instant adjustment. Acquisition, Analysis, and Sorting.
Star Wars: Knight Errant: Influx
Index Sorting. Unique properties revealed with polarization-sensitive detectors. Prochlorococcus and polystyrene bead scatter using the Small Particle Option.
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Hierarchical gating strategy. Four-way sorting. Analysis of aquatic samples.
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Measurement of macrophage cell line stimulation using GFP. Cy dyes are subject to proprietary rights of Amersham Biosciences Corp and Carnegie Mellon University and are made and sold under license from Amersham Biosciences Corp only for research and in vitro diagnostic use. Any other use requires a commercial sublicense from Amersham Biosciences Corp. Please Select. Clinical instruments Research instruments Clinical cell analyzers Clinical sample prep Clinical software. We examined cytospin slides of nasal washings obtained before and hourly for 11 h after nasal antigen challenge in 10 asymptomatic allergic subjects with a history of seasonal rhinitis and 5 normal, nonallergic subjects.
Allergic subjects received oral prednisone 20 mg 3 times a day or placebo in a random, double-blind crossover manner for 2 days before each of 2 challenges 1 month apart. On placebo days, a mixed cell influx occurred in allergic subjects during the late response that was fold greater than the cell influx in the nonallergic control subjects p less than 0. During the first 3 h after antigen challenge, eosinophils p less than 0. During the late phase 4 to 11 h , neutrophils, eosinophils, and mononuclear cells were all increased.