CRG/UPF Flow Cytometry Unit

CORE FACILITIES

HEAD OF THE UNIT:
Òscar Fornas (UPF)

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

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 its staff’s expertise fully cover both the wide range of conventional flow cytometry applications and the more 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, development and dissemination

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

2017
Analyzers usage (h) 4.425
Cell Sorting usage (h) 2.190
Total usage (h) 6.615
Total invoicing (€) 177.052

New collaborations have been stablished on chromosome sorting for chromosome sequencing in basic research and agro-sciences (i.e. with Dr. Tomàs Marquès, for Chromosome Sorting for chromosome sequencing without amplification; and with Dr. Rosa Figueroa, Instituto Español de Oceanografía, Vigo, Spain, for Sorting of chromosomes in Dinoflagellates and unicellular sea organisms).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.

Other ongoing collaborations are with Dr. Toni Gabaldón, from CRG, for Single-cell genomics and transcriptomics of mouth’s microbioma; and with Dr. Jordi Garcia-Mas IRTA-CRAG, UAB, Bellaterra, Spain,  for Sorting of pollen’s microspores for RNAseq and Miseq.

The head of the Unit was involved in developing a novel application of single-virus genomics sorting that was finally been published in Nature Communications:

Martinez-Hernandez F, Fornas O, Lluesma Gomez M, Bolduc B, de la Cruz Peña MJ, Martínez JM, Anton J, Gasol JM, Rosselli R, Rodriguez-Valera F, Sullivan MB, Acinas SG, Martinez-Garcia M.
‘Single-virus genomics reveals hidden cosmopolitan and abundant viruses.’
Nat Commun, 8:15892. doi: 10.1038/ncomms15892 (2017).

The Head of the Unit also participated in different training or teaching courses: At UPF and CRG masters courses and flow cytometry courses at other institutions in Barcelona, Badalona and Madrid.

Has also hosted as a training workshop to advance in multicolour panel design for multiparametric flow cytometry assays in collaboration with Becton Dickinson. He was also invited as speaker to talk about new developed methodology of identification and isolation of exosomes by flow cytometry at Erasmus University (Rotterdam); at SIC (Sociedad Iberica de Citomeria) for a plenary session; at GENYO in Granada; and at IMIBIC in Cordoba.