FACS Expertise Unit

Headed by: Jochen Lamote

Flow Cytometry is a widely-used field to characterize the expression of proteins on a single-cell level. Proteins in the cell are either labeled with antibodies conjugated to a fluorophore or with fluorescent dyes. Depending on the instrument, several proteins can be simultaneously detected on a single cell, including a microscopic image to assess localization of your protein of interest in the cell. Next to characterizing the cells with analyzers, cells can be sorted based on their protein expression or simply based on morphological parameters to purify the cells of interest for a wide range of downstream applications, including single cell RNA-sequencing, mass spectrometry, functional assays, and others.

The CCB FACS Expertise Center offers technical and scientific support on FACS-related experiments. The platforms are high throughput and the sorting and analysis software are relatively easy-to-use.


Mission

  • Generate high quality data, using the right technical controls, excluding variability, appropriate panel design.
  • Highly flexible, support needs of the projects and adjust to continuously changing timelines, access is granted 24/7 after training.
  • Combining relatively easy accessible equipment with a relatively high flexibility so in-house trained people can use the equipment independently.
  • Assistance for cell sorting, data-acquisition and data-analysis
  • Providing training for cell sorting and Multicolor panel design
  • Driving innovation of downstream applications (e.g. transcriptomics) by overcoming technical difficulties related to cell sorting.

In-house Equipment

  • BD Fortessa X-20

The BD Fortessa X-20 is an analyzer with a robust reputation over the years. The device is relatively easy to use. Especially users with pre-existing knowledge on FACS DIVA (e.g. BD Canto, BD Aria) will have no issues to use the instrument. Training – based on the FACS experience level - is provided. The Fortessa features 20 parameters (18-colors + 2 physical parameters) which are customizable based on user request.

Standard Configuration:

    • UltraViolet laser: BUV395, BUV496, BUV563, BUV805 (optional: BUV737)
    • Violet laser: BV421, BV510, BV605, BV650, BV711, BV786
    • Blue laser: FITC, PerCP-Cy5.5
    • YellowGreen laser: PE, PE-CF594, PE-Cy7 (optional: PE-Cy5; PE-Cy5.5)
    • Red laser: APC, AF700, APC-Cy7
       Fortessa

       

  • Sony MA900

The Sony MA900 is an operator-friendly Cell Sorter.The system is easy and straightforward to use, which results in some compromises, like collinearity of the lasers. The system runs on disposable sorting chips, which eliminates the built of contaminants in the Flow Cell in conventional cell sorters. This instrument has a low threshold for people who have no pre-existing experience with cell sorting.

Configuration:

    • Violet + Red laser:
      • BV421, BV510, BV570, BV605, BV650 or APC, BV711 or AF700, BV786 or APC-Cy7
    • Blue + YellowGreen laser:
      • FITC, PE, PE-CF594, PERCP-Cy5.5 or PE-Cy5.5, PE-Cy7
         Sony