UNIT HEAD:  Òscar Fornas (UPF-CRG)

OPERATORS: Erika Ramírez (CRG), Eva Julià (IMIM), Àlex Bote (CRG)

SUMMARY

The CRG/UPF joint Flow Cytometry Unit provides PRBB researchers with technical expertise and training to access state-of-the-art instrumentation, as well as technical and scientific advice to develop efficient and reliable flow cytometric assays with the highest quality control standards and productivity. The unit’s instrumentation and staff’s expertise fully cover the wide range of conventional flow cytometry applications and most advanced ones. New applications are developed and implemented to respond to the facility’s needs or by user demand, thus maintaining the unit at the forefront of flow cytometry. The current personnel cover the entire demand for cell sorting and user assistance needs in experimental design, advice and training.

EQUIPMENT

The CRG/UPF Flow Cytometry Unit is one of the best-equipped facilities in Europe, boasting some of the most advanced technology to support all flow cytometry applications.

The facility’s equipment is currently comprised of six analysers and two cell sorters, as listed below:

Instrument

Laser (nm)

Detectors

Situation

BD FACScalibur analyser

488 633

FSC/SSC + 4PMT

Active

BD FACSCanto analyser

488 633

FSC/SSC + 6PMT

Active

BD LSR II analyser

325 407 488 633

FSC/SSC + 10PMT

Active

BD LSR II analyser

325 488 633

FSC/SSC + 8PMT

Active

BD LSRFortessa analyser

407 488 561 633

FSC/SSC + 14PMT

Active

SONY SP6800 spectra analyser

407 488 640

FSC/SSC + 34 ch

Active

BD FACSAria II SORP cell sorter

325 407 488 561 633

FSC/SSC + 18PMT

Active

BD Influx cell sorter

325 457 488 561 633

FSC/SSC + 14PMT

Active

ACTIVITY

The activity in the facility can be described in terms of hours of analysers and cell sorting usage, and translated into invoicing. Both indicators are shown in table below:

2018

Analyzers usage (h)

3.613

Cell Sorting usage (h)

2.511

Total usage (h)

6.124

Additionally, training, teaching, technology or methodology development in collaborations are essential to our philosophy of facility management, to properly assist and help users driving flow cytometry in their fields.

DEVELOPMENT

The Flow Cytometry Unit promotes development of new applications both based on the research community needs and directly through collaborations with researchers.

Main highlights regarding these collaborations are:

  • Bivariate Flow Karyotyping.
  • Nano-particle detection and isolation (i.e. viruses and exosomes).
  • Single-particle isolation for RNAseq, DNAseq, etc, for basic research and agro-sciences in different models:
    • Mammalian cells.
    • Viruses.
    • Bacteria.
    • Global microbiome.
    • Pollen.

TRAINING AND TEACHING

To access the state-of-the-art technology, technical training is essential. The facility puts a lot of efforts in providing the necessary training to users.

  • Internally, the facility staff has provided more than 300 hours of individual training to users.
    • 235 hours to hands-on training (programmed training).
    • 73 hours to theoretical training.
  • As every year, the facility hosted again a workshop to advance in multicolour panel design for multiparametric flow cytometry assays, in collaboration with Becton Dickinson (this time addressed to internal users).

The Head of the Unit also participated in different training or teaching courses at UPF and CRG master’s courses and flow cytometry courses at other institutions in the Barcelona area.

SELECTED PUBLICATIONS

Martinez-Hernandez F, Fornas Ò, Lluesma Gomez M, Garcia-Heredia I, Maestre-Carballa L, López-Pérez M, Haro-Moreno JM, Rodriguez-Valera F, Martinez-Garcia M.
Single-cell genomics uncover Pelagibacter as the putative host of the extremely abundant uncultured 37-F6 viral population in the ocean.
ISME J, 18:10, 2018.

de la Cruz Peña MJ, Martinez-Hernandez F, Garcia-Heredia I, Lluesma Gomez M, Fornas Ò, Martinez-Garcia M.
Deciphering the Human Virome with Single-Virus Genomics and Metagenomics.
Viruses, 10(3). pii: E113, 2018.