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BD LSRFortessa Special Order Research Product 5 Laser Cell Analyzer Flow Cytometer

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Includes HTS, HP Z220 Workstation with FACSDiva Software Version 7

Technical Specifications:

Excitation Optics

  • Excitation Optical Platform:
    • The special order BD LSRFortessa optical layout allows for up to five lasers.
  • Laser Wavelengths and Laser Powers:
    • Blue Green: 100 mW with 2 PMT detectors
      • YFP
      • mOrange
    • Violet Laser 405 nm: 100 mW with 2 PMT detectors
      • Pacific Blue
      • AmCyan
      • Pacific Orange
      • LSS mOrange
    • Blue 488 nm: 200 mW with 5 PMT detectors plus side scatter
      • FSC
      • SSC
      • FITC
      • PE
      • PerCP-Cy5.5
    • Yellow Green 561 nm: 100 mW with 5 PMT detectors
      • PE-YG
      • PE-Texas Red
      • PE-CY5
      • PE-Cy5.5
      • PE-Cy7
    • Red Laser 637 nm: 100 mW with 3 PMT detectors
      • APC
      • Alexa Flour 700
  • Flow Cell Design:
    • Rectangular Quartz Cuvette: Internal cross-section, 430 x 180 μm
    • Fixed optical assembly of the lasers, with up to 5 spatially-separated laser beams.

Emission Optics

  • Optical Coupling:
    • The quartz cuvette flow cell is gel coupled by refractive index-matching optical gel to the fluorescence objective lens (1.2 NA) for optimal collection efficiency.
  • Forward Scatter Detection:
    • Photodiode detector with a 488/10 bandpass filter
  • Side Scatter Detector:
    • Photomultiplier tube (PMT) with a 488/10 bandpass filter
  • Emission Optical Design:
    • The special order BD LSRFortessa cell analyzer uses BD’s patented octagon and trigon detection system. Using reflective optics and fiber-coupled PMTs ensures more efficient light collection than that obtained from transmission optics.


  • Fluorescence Sensitivity:
    • FITC-A: 80 molecules of equivalent soluble fluorochrome (MESF-FITC)
    • PE-A: 30 molecules of equivalent soluble fluorochrome (MESF-PE)
    • Measurements performed using SPHERO Rainbow Calibration Particles RCP-30-5A
  • Fluorescence Resolution:
    • Coefficient of variation PI: area of <3%, full G0/G1 peak for chicken erythrocyte nuclei (CEN) stained with propidium iodide (PI)
  • Fluorescence Linearity:
    • Doublet/singlet ratio of 1.95–2.05 for CEN stained with PI and excited with the 488-nm blue laser
  • Side Scatter Sensitivity:
    • Achieves separation of 0.5-µm beads from noise in side scatter.
  • Forward and Side Scatter Resolution:
    • Scatter performance is optimized for resolution of lymphocytes, monocytes, and granulocytes.
  • Forward Scatter PMT Option:
    • A forward scatter PMT upgrade is available for small particle detection through the special order program.
  • Data Acquisition Rate:
    • Theoretical maximum electronic data collection of 70,000 events/second.
    • Typical operation at 20,000 events/second which requires a sample concentration of 2 x 107 cells/mL at an instrument flow rate of 60 μL/min


  • Sample Flow Rates:
    • Front key panel provides three modes: RUN, STNDBY, and PRIME
    • Continuously adjustable flow rate, plus three preset flow rates:
      • LO = 12 μL/min
      • MED = 35 μL/min
      • HI = 60 μL/min
  • Standard Fluidic Reservoirs:
    • One 8-L sheath and 10-L waste container provided.

Performance without peer, choice without compromise
The BD LSRFortessa cell analyzer offers the ultimate in choice for flow cytometry, providing power, performance, and consistency. Designed to be affordable and expandable, the BD LSRFortessa has the flexibility to support the expanding needs of multicolor flow cytometry assays.

Flexibility in a Small Space
An affordable choice to fit most flow cytometry analyzer needs, the BD LSRFortessa system can be configured with up to 7 lasers*—blue, red, violet, UV and yellow-green. This flexibility in laser wavelengths allows assay design to be optimized using the latest fluorescent dyes and substrates. The instrument can accommodate the detection of up to 18 colors simultaneously with a defined set of optical filters that meet or exceed the majority of today’s assay requirements. The BD LSRFortessa is an innovative and proven platform for multicolor analysis.

The BD LSRFortessa puts the power of the proven BD™ LSR platform into a compact footprint. It can easily fit on the benchtop for cost effective space use.

This space efficiency is even more important for labs with limited space or where space cost is at a premium. In addition to the reduced size, design enhancements provide easier access to bandpass filters and mirrors, simplifying changes to experimental setup.

Superior Performance
Many innovations are incorporated into the BD LSRFortessa product line. The heart of the cytometer, the fluidics system features a true fixed alignment flow cell that is gel-coupled to the collection optics to maximize detector signal

Fluidics System
The fluidics system is pressure driven. Hydrodynamic focusing forces sample cells through the cuvette flow cell, where they are interrogated. The flow cell is in fixed alignment with the laser and gel-coupled to the collection optics. This design ensures that the laser is precisely focused on the sample stream and the maximum amount of emitted light can be collected for added sensitivity in multicolor applications. Fixed alignment also minimizes startup time, improves experiment-to-experiment reproducibility, and enables automated daily quality control.

The sheath container (8 L) and waste container (10 L) are outside the cytometer, and positioned on the floor for easier access.

Fluidic sensors maintain constant pressure, while a fluidics monitoring system warns when sheath fluid is low, empty, or when the waste container is full.

Maximum Signal, Minimum Crosstalk
The patented collection optics are yet another design innovation. Arranged in octagon- and trigon-shaped optical pathways, their novel design efficiently maximizes signal detection and increases sensitivity and resolution. This allows researchers to identify cells, especially dim and rare cell populations, optimizing multicolor assays and panel design for superior results.

Optics System
The optics system consists of laser excitation optics that illuminate cells in the sample, and collection optics that direct light scatter and fluorescence signals through spectral filters to detectors. Innovative designs for both the excitation optics and collection optics in BD LSRFortessa systems reduce excitation losses and optimize collection efficiency for increased sensitivity and resolution.

Excitation Optics
The excitation optics consist of multiple fixed wavelength lasers, beam shaping optics, and individual pinholes which result in spatially separated beam spots.

A final lens focuses the laser light into the gel-coupled cuvette flow cell. Since the optical pathway and the sample core stream are fixed, alignment is constant from day to day and from experiment to experiment.

Collection Optics
Emitted light from the gel-coupled cuvette is delivered by fiber optics to the detector arrays. The collection optics are set up in patented octagon- and trigon-shaped optical pathways that maximize signal detection resulting from each laser illuminated beam spot. Bandpass filters in front of each PMT allow spectral selection of the collected wavelengths. Importantly, this arrangement allows filter and mirror changes within the optical array to be made easily and requires no additional alignment for maximum signal strength.

This design is based on the principle that light reflection is more efficient than light transmission. Emitted light travels to each PMT detector via reflection and is transmitted through only two pieces of glass to reach the detector. Therefore, more colors can be detected with minimum light loss.

Greater Consistency
BD FACSDiva software controls the efficient setup, acquisition, and analysis of flow cytometry data from the BD LSRFortessa workstation. The software is common across BD FACS™ instrument families, including the BD FACSCanto™ cell analyzer and BD FACSAria™ cell sorter systems. This affords researchers greater application flexibility allowing them to easily move assays from one platform to another.

Cytometer Setup and Tracking
The Cytometer Setup and Tracking (CS&T) feature of BD FACSDiva software establishes baseline settings and adjusts for instrument variability. The software reduces operator error, and ensures consistency of results by setting the signal time delay across the multiple laser beams and optimizing PMT voltages. Application specific settings can also be set, allowing for rapid setup and performance of routine experiments in a more consistent manner. Quality control (QC) tracking capabilities in the software measure instrument settings and report on performance. Levey-Jennings plots help users understand instrument performance and identify maintenance issues.

Acquisition and Analysis
BD FACSDiva software enables researchers to preview and record data from multiple samples with an automated acquisition process. Acquisition templates, user-definable experiment layouts, and simple compensation procedures are also managed by the software to further facilitate acquisition.

BD High Throughput Sampler Option
To improve experimental workflow, the optional BD High Throughput Sampler (HTS) provides rapid, fully automated sample acquisition from 96- and 384-well microtiter plates. In high-throughput mode, the HTS option can speed through a 96-well plate in fewer than 15 minutes with less than 0.5% sample carryover from one well to the next. Low carryover is essential in research applications to ensure sample purity and data integrity.

Fast acquisition speed is achieved by synchronizing two high-precision pumps for sample mixing, sample injection, and probe washing. Standard throughput mode can be selected for acquisition of larger sample volumes.

Special Order Products
The BD special order research program allows customers to configure BD flow cytometers and cell sorters to fit precise research and assay needs. Tailored to the special needs of research at the leading edge of biomedical discovery, the program offers a wide range of choices to help researchers create the ultimate customized instrument for their requirements.

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LSR Fortessa SORP

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