Image-Enabled and Spectral Cell Sorter

BD FACSDiscover S8

The BD FACSDiscover S8 Image-Enabled Spectral Cell Sorter is the first instrument that is capable of cell sorting based on imaging data. Patented CellView™️ high-speed imaging technology facilitates spatially resolved signal measurements without a camera by using the optical and electronic components available on flow cytometers.

In addition to image-enabled sorting, the FACSDiscover S8 incorporates a spectral detection system based on avalanche photodiodes (ADPs) measuring fluorescence from 5 laser lines for true high parameter sorting.

Here are some of the unique applications made possible by the FACSDiscover S8.

1. Image-Enabled Sorting: Utilize quantified imaging data (“features”) in your sort logic. Imaging features can be plotted and gated like traditional flow cytometry data and used for sorting
   • Differentiation of punctate from diffuse fluorescence signal
   • Detection of the degree of colocalization of two fluorescence signals
   • Measurement of the distance between two fluorescence signals
   • Detection of cell-cell interactions
   • Enhanced doublet exclusion
   • Sorting cells based on cell size
2. High parameter spectral cell sorting of panels of at least 50 colors*.
3. Six-way sorting to isolate up to six populations simultaneously from a sample.
4. Single cell deposition directly onto plates, including 384-well plates.
5. Index sorting of both imaging and flow cytometry data when sorting into plates.
6. 85, 100, and 130 μm nozzles to accommodate a variety of sample types.
7. Biosafety cabinet for sorting of primary human tissue.

* Konecny AJ, Mage PL, Tyznik AJ, Prlic M, Mair F. OMIP-102: 50-color phenotyping of the human immune system with in-depth assessment of T cells and dendritic cells. Cytometry A. 2024 Jun;105(6):430-436. doi: 10.1002/cyto.a.24841. Epub 2024 Apr 18. PMID: 38634730; PMCID: PMC11178442.

Image-Enabled Sorting

Imaging Specifications:

1. Six Imaging Channels:

   • 3 morphological/non-fluorescence images (forward scatter, side scatter, and light loss)

   • 3 fluorescence images

2. Magnification: ~15x
3. Resolution: 1.5 μm²/pixel
4. Field of View: 60 μm

Imaging Features

Imaging features permit quantification of imaging measurements in experimentally useful ways. These quantified features can be plotted like flow cytometry data and used for sorting. The following features are currently available on the FACSDiscover S8:

• Light Loss
• Forward Scatter
• Side Scatter
• Correlation
• Delta Center of Mass
• Diffusivity
• Eccentricity
• Max Intensity
• Radial Moment
• Center of Mass (X)
• Center of Mass (Y)
• Long Moment
• Short Moment
• Size
• Total Intensity

Imaging Configuration

Spectral Cell Sorting

The FACSDiscover S8 is equipped with a state-of-the-art spectral detection system coupled with BD patented SpectralFX™️ electronics and algorithms.

Highlights of SpectralFX™️ include:

1. Full spectrum optics and optimized hardware design with 78 solid state detectors and 5 lasers for all the classical benefits of spectral flow cytometry.
2. System aware unmixing algorithm adapts to your sample and instrument in real time to manage spread.
3. Next generation QC system uses LEDs and beads to measure noise, perform gain calibration and provide real time noise/signal hardware information.
4. Guided workflow allows users to learn quickly and use best practices in experimental setup.

The FACSDiscover S8’s SpectralFX™️ technology is equipped with the following hardware components for spectral detection.

1. Five spatially separated excitation lasers
   • 349 nm (UV)
   • 405 nm (violet)
   • 488 nm (blue)
   • 561 nm (yellow green)
   • 637 nm (red)
2. 78 fluorescence avalanche photodiode (ADP) photodetectors

^*488 nm laser is split between the spectral detection system and the CellView™ imaging system.