BD Influx™

The flexible BD Influx™ cell sorter easily adapts to a researcher’s application or environmental requirements. A modular architecture, hands-on controls, support for up to ten lasers, exchangeable detector options, high throughput, and six-way sorting make the system highly configurable to precise needs of advanced research.

In addition, BD FACS™ Sortware sorter software gives researchers comprehensive control, from configuration and compensation setup to acquisition, sorting, and analysis.

Innovative Fluidics Design

The BD Influx™ fluidics system features a unique acoustical coupling in the nozzle assembly to reliably create droplets for sorting, while ensuring low shear stress to optimize cell viability, even at high pressures. To support aseptic sorting, the fluidics path is easily removed and replaced with the optional disposable fluidics kit.

Nozzle assembly and nozzle sizes

The core of the BD Influx system is the nozzle assembly, which features an optimized acoustical coupling, enabling the most efficient droplet formation. This unique design minimizes noise from the droplet formation process and results in high droplet formation rates at relatively low sheath pressures.

In addition, the form of the nozzle assembly enables cells to accelerate smoothly to the laser intercept, increasing the viability and functionality of even the most fragile cells. A variety of nozzle sizes are available to allow researchers to easily adapt the system to different cell or particle types.

Sample station and fluidics console

A sample station and fluidics console control sample flow, sample line backflush, boost, and differential pressure. Bubble detectors prevent air bubbles from reaching the nozzle by stopping flow when the sample tube is empty.

Stainless steel tanks

Stainless steel pressurized sheath and waste tanks allow researchers to adapt cleaning procedures to match their application needs.

Exchangeable fluidics

Parsley 1 Image - Fluidics Nozzle

An important concern in cell sorting is cross-contamination, either between samples or with foreign biological agents such as bacteria, viruses, and DNA. To mitigate the risks of cross-contamination, researchers can replace the sample line in a matter of minutes. The sample line extends into the nozzle tip to minimize contact between cells and non-replaceable parts. For rigorous control, the complete fluidics system, from sheath tank to nozzle tip, can be exchanged. The unique design of the BD Influx fluidics system allows for rapid exchange of all parts that potentially contact cells, addressing cross-contamination concerns.

Pressurized stainless steel sheath and waste tanks

Intuitive Alignment

The optical system that directs lasers to the interrogation point is easy to use, intuitive, and adaptable. Up to 10 laser paths can be configured. BD is constantly expanding the choice of lasers available for the BD Influx™ system. Check the current technical specifications for the most up-to-date information.

The 10 laser paths and 7-pinhole optical collection system support 24 parameters simultaneously and 24 x 24 compensation. Systems with up to 5 lasers support 16 simultaneous parameters and 16 x 16 compensation.

For each laser, light is shaped by individual optics, and final alignment and focusing are achieved using a dedicated lens that steers light to the stream at different angles. For the ten-laser system, the laser paths are collinear through the steering optics and focusing lens, but intersect the stream at seven spatially separated points. Fluorescence is collected through a high NA lens and focused on individual mirrored pinholes, reducing crosstalk between lasers. The modular design of the BD Influx optical system makes it easy to upgrade lasers as research requirements change. In addition, the variety of available lasers is constantly growing to keep pace with the needs of advanced research.

The patented optical design uses a special pinhole camera to image the stream and the pinholes simultaneously, simplifying optical alignment. With the help of the pinhole camera, near optimal alignment can be achieved within seconds without using beads.

Optics Features - Focus Lens

Pinhole camera view

Schematic representation of collection lens, pinhole camera, and mirrored pinholes

Individual laser positioners

Flexible Sorting

The BD Influx™ system features two-, four-, and six-way sorting, as well as plate sorting. The BD Influx electronics support high-speed sorting, and BD FACS™ Accudrop technology helps simplify sort optimization.

In addition, new BD FACS™ Sortware sorter software facilitates sorting QC by allowing users to learn about how sort parameters impact results, to optimize research.

Multi-way sorting

Isolating multiple populations at once saves time and makes efficient use of sample. Whether sorting one subpopulation or six, the BD Influx sorter can be quickly configured to best fit researchers’ application requirements. A wide range of standard collection options supports two-, four-, and six-way sorting. Tube holders include sizes from microtubes, to 12 x 75-mm tubes, to 15-mL tubes.

Electronics

The BD Influx system uses parallel electronics to reach a throughput rate of 200,000 events per second, independent of the number of parameters. For high sensitivity and low noise, the BD Influx system uses analog electronics in the first stage of signal amplification. The output of the analog peak and hold circuitry is digitized using 16-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. When these factors are taken into account, the yield can be calculated by using Poisson’s statistics. The BD Influx system will reach >80% of Poisson’s expected yield with properly prepared samples.

Accudrop technology simplifies drop-delay determination

BD FACS Accudrop technology simplifies setup and eliminates manual calculations normally required. 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 drop-delay value can be adjusted while viewing BD FACS™ Accudrop beads, which are illuminated in the center and side sort streams. The researcher can identify the precise drop-delay value without a fluorescence microscope or manual calculations.

Adaptability

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.

Sorting Features - Small Particle no polarization

Small Particle Option

Optics Features - Small Particle Option

Small Particle Option with polarization

Polarization

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.

Small Particle Option

The Small Particle Option on the BD Influx system improves forward scatter detection. 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 (0.1-micron filtered) lowers the intrinsic noise level, which enables measurement of particles as small as 200 nm.

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.

Computerized Cell Deposition Unit

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. Single-cell cloning or sorting for single-cell PCR or single-cell imaging techniques is possible with the standard CCDU.

Make exploratory cloning more efficient with index sorting

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 96-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. Each sorted event in the file is “indexed” one-by-one according to the X and Y coordinates of the sort collection device. Post-sorting results can be precisely traced back to the flow characteristics of the specific cell or combinations of cells sorted.

Flexible collection formats

Cell Deposition Unit - Large

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 384-, 96-, 24-, and 6-well plates, coarse and fine calibration slides, and 4 x 12 slides.

Custom Cabinet Meets Biosafety Standards

A biological safety cabinet (BSC) designed specifically for the BD Influx™ cell sorter by The Baker Company, measuring 86.2 x 53.5 x 94 in. (219.0 x 136 x 239 cm) is available as an option. Biological safety in flow cytometry is an emerging requirement for core laboratories concerned about the potential accidental exposure of operators to biological samples.

Protecting personnel, products, and the environment

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.

Multiple levels of control to match application and environmental needs

Verified to meet biosafety standards

The Baker Company has verified that the BSC designed specifically for the BD Influx sorter meets personnel and product protection standards for a Class II Type A2 biosafety cabinet and the National Sanitation Foundation International Standard 49. 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.

Airflow control

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. Aerosols from the BD Influx sort chamber are directly evacuated into the high efficiency particulate air (HEPA) filters of the BSC, eliminating the need for an aerosol management option. 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 99.97% of particles or equivalent with a diameter of 0.3 μm.

BD Influx

BD FACS Sortware: from Acquisition to Sorting and Analysis

BD FACS™ Sortware sorter software provides comprehensive control of the cell sorter from configuration and compensation setup to acquisition, sorting, and analysis, making it easy to customize the instrument for specific applications.

BD FACS Sortware fundamentally improves the way cytometer software supports cell sorting. 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.

To support analysis, BD FACS Sortware lets researchers visualize data from experiments in a variety of rich output formats. Output formats include histograms, overlay histograms, and dot, density, and contour plots—all with linear, log, or biexponential (Logicle) scaling. Support for standard software protocols allows researchers to export both graphical data and text into standard productivity software (such as Microsoft® Office®) for presentation or publication. Results can also be exported and imported as FCS data files for other software applications.

Intuitive workflow

BD FACS Sortware has a simple, intuitive workflow, and its familiar Microsoft® Windows® interface allows researchers to focus on their experiment rather than on commands and dialog boxes. 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.

Cytometer

Acquisition, Analysis, and Sorting

Compensation

Sorting

Sorting

Index Sorting

Reagents

Featured Product

  • BD Horizon Brilliant™ Polymer Dyes
    For reagent selection in general, "bright is better." For cells that have few receptors on the surface, bright reagents are essential in resolving these dim cells from others in a sample. BD Horizon Brilliant™ polymer dyes are four to ten times brighter than conventional dyes, with equivalent background—a breakthrough in the field.

BD FACSAria™ Fusion Violet Laser (405 nm)

BD FACSAria Fusion Blue Laser (488 nm)

BD FACSAria Fusion Yellow-Green Laser (561 nm)

BD FACSAria Fusion Red Laser (640 nm)

Buffers

BD FACSAria Fusion UV Laser (355 nm)


Unique properties revealed with polarization-sensitive detectors

Prochlorococcus and polystyrene bead scatter using the Small Particle Option

Hierarchical gating strategy

Four-way sorting


Real-time viewing

Discrimination of small particles can be further improved by measuring fluorescence


Analysis of aquatic samples

Measurement of macrophage cell line stimulation using GFP


For Research Use Only. Not for use in diagnostic or therapeutic procedures.

Microsoft, Windows, and Office are registered trademarks of Microsoft Corporation.

Class 1 Laser Product.

Alexa Fluor® and Texas Red® are registered trademarks, and Pacific Blue™ is a trademark, of Molecular Probes, Inc.

Cy™ is a trademark of Amersham Biosciences Corp. 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. 800 Centennial Avenue, Piscataway, NJ 08855-1327, USA.