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Applications & Techniques

Overview
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BD SpectralFX™ Technology combines an optimized hardware design for full-spectrum optical detection with a system-aware unmixing algorithm to maximize the palette of colors that can be used for flow cytometry with optimal spectral resolution and sensitivity. BD SpectralFX™ Technology is paired with a next-generation setup and quality control (QC) system and a guided software workflow that helps to automate detector setup for consistency and performance and to simplify the spectral workflow.

 

The benefits of BD SpectralFX™ Technology and  BD CellView™ Image Technology are integrated into the BD FACSDiscover™ S8 Cell Sorter, a real-time imaging and spectral flow cytometer. This combination enables the measurement of an unprecedented number of features per cell, expanding the power of cell analysis and sorting, and empowering scientists to address previously impossible-to answer questions.

Optimized Optical and Hardware Design

BD SpectralFX™ Technology uses a modular optical architecture that minimizes optical losses to allow higher detector count. The technology allows expansion of panel size and fluorochrome selection flexibility with 78 fluorescent detectors across five lasers with algorithmically optimized bandpass filters.

 

Excitation Optics:

  • 349 nm
  • 405 nm
  • 488 nm*
  • 561 nm
  • 637 nm
 
*488-nm laser is optically divided to support signal detection with BD CellView™ Image Technology
performance1

 

 

Overview of BD FACSDiscover S8 Optical Layout

Simplified overview of the optical layout for the BD FACSDiscover™ S8 Cell Sorter.

The BD FACSDiscover™ S8 Cell Sorter is equipped with five spatially separate lasers and corresponding spectral detector arrays.

Next-Generation Setup and QC

Next-generation setup and QC automates detector setup for consistency and performance. An on-board LED is used to perform detector gain calibration and characterize detector sensitivity as part of daily setup, enabling simple and consistent detector gain control across all detectors. In addition, a broad-spectrum setup bead is used to track full-system performance and to ensure consistent measurement results over time through MFI tracking.

 

Default detector gain settings are available for each experiment to provide a balance between detector sensitivity and dynamic range, eliminating the need to manually adjust voltages for every experiment.

Next Generation Setup and QC

System-Aware Unmixing Algorithm

Many factors, such as fluorochrome spectral overlap, electronic noise, and optical background, contribute to spread in unmixed data leading to loss of biological resolution. BD SpectralFX™ Technology uses a system-aware unmixing algorithm that adapts the unmixing process to system noise, sample quality and detector gains to help manage spillover spreading in real time.

 

standard and System-Aware Unmixing

 

System-aware unmixing manages spread across a wide range of detector gain settings using LED-based detector noise calibration and real-time noise estimates for each fluorescence detector, helping manage the spread of unmixed population across the spectrum.

 

BD SpectralFX™ Technology combines full-spectrum optics, next generation QC and system-aware spectral unmixing that manages spread by adapting to instrument performance and sample conditions in real time for sort-compatible unmixing.

Guided Workflow

BD FACSChorus™ Software uses a guided workflow to support best practices while staying flexible for new applications. A step-by-step workflow is provided with all the necessary steps to successfully perform an experiment:

FACSChorus Guided Workflow
FACSChorus Guided Workflow Part 1
  • Design Experiment: Select markers and fluorochromes for the experiment.
  • Select Imaging Features
  • Adjust Gains: New detector control interface helps adapt gain settings for new samples by easily seeing off-scale channels.
FACSChorus Guided Workflow Part 2
  • Set Up Single-Stain Controls: Record unstained samples, record single-stain controls, define appropriate scatter gates and define your positive and matching negative populations.
  • View Data: Collect fully-stained samples and visualize unmixed results.
  • Set Up Sort
  • Sort
  • View Reports
Sample Data
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Six-Way Sorting of Deep Immunophenotyping Panel

This 38-color spectral panel characterizes and sorts deep lineages of T cell and NK cell subsets. The panel includes BD Horizon RealYellow™ and BD Horizon RealBlue™ dyes, engineered to be bright, clean and laser specific to work in tandem on the BD FACSDiscover™ S8 Cell Sorter for high-parameter spectral analysis to be able to reveal complex biological information.

38-color spectral panel characterizes and sorts deep lineages of T cell and NK cell subsets
38-color spectral panel characterizes and sorts deep lineages of T cell and NK cell subsets

Six-way sorting of deep immunophenotyping panel. This 38-color spectral panel characterizes and sorts deep lineages of T cell and NK cell subsets.

BD Solutions
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Combining BD SpectralFX™ Technology with BD CellView™ Image Technology

The BD FACSDiscover™ S8 Cell Sorter with BD CellView Image Technology and BD SpectralFX Technology, the first spectral flow cytometer sorter with sort-capable image analysis, expands the power of cell analysis and sorting to new dimensions by combining spectral flow cytometry with real-time spatial and morphological insights—empowering scientists to address previously impossible-to-answer questions.

 

  • Obtain insights on cell populations and characteristics that can be visually confirmed in real time during analysis and sorting.
  • Enhance spectral flow cytometry with spatial and morphological insights to interrogate and sort cell types that previously could not be identified or isolated.
  • Simplify your workflow with built-in visual inspection capabilities, more flexible panel design and automated features.
  • Combine single cell imaging, spectral flow and downstream analysis to create a comprehensive profile of complex diseases.
Learn More
 

BD flow cytometers are Class I Laser Products.

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