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 instrumentation and staff expertise fully cover the wide range of conventional flow cytometry applications as well as most advanced ones. New ones are developed and implemented to respond to the facility’s needs or by user demand, maintaining the unit at the forefront of flow cytometry. The current personnel meet the entire demand for cell sorting as well as users’ assistance needs in experimental design, advice and training..
Equipment
The CRG/UPF Flow Cytometry Unit is one of the best-equipped units in Europe with some of the most advanced technology to support all flow cytometry applications.
Currently six analysers and two cell sorters make up the facility equipment as follows:
| Instrument | Laser (nm) | Detectors | Situation |
|---|---|---|---|
| BD FACScan analyser | 488 | FSC/SSC + 3PMT | Active |
| 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 LSRFortessa analyser | 407 488 561 633 | FSC/SSC + 14PMT | Active |
| SONY SP6800 analyser * | 407 488 640 | FCS/SSC + 30PMT | 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 |
*A SONY SP6800 spectral analyser has recently been purchased by UPF providing the newest Flow Cytometry technology. This is a new flow cytometry technology with high cell analysis capabilities for multiparametric cell analysis as well as new flow cytometry generation.
Additionally, a High Throughput Sampler (HTS) for a sample multiloader system has also been purchased by UPF and installed on the BD Fortessa analyser to cover high-throughput sample analysis required for screening hundreds of samples in a short period of time.
A biosafety cabinet has also been purchased to fit the FACSAria cell sorter in it, advancing in operator biosafety and being equipped to offer the possibility to handle cell sorting procedures up to a BSL2-like level.
Activity, development and dissemination
The activity in the facility can be described by hours of analysers and cell sorting usage. Both of them are key activity indicators. While the analysers have decreased by 8% (from 4,129 in 2014 to 3,802 hours in 2015), the cell sorting usage has increased by 12.7%, from 2,365 in 2014 to 2,666 hours in 2015. Globally, in 2015 activity increased by 1.5% over 2014.
After the implementation of high-resolution applications to advance in the genomic field during 2014 (i.e. Flow Karyotyping for chromosome sorting), external researchers contacted us to set up hot new topics on flow cytometry applications and perform nano-particle isolation and sorting (i.e. virus and exosomes). Both of them are pending publication. When published, they are sure to be appreciated by the scientific community, as both are valuable tools in biomedicine research. For years, the scientific community has focused its efforts on developing them and we are currently the only facility capable of carrying out those applications in a reliable way. This will motivate more researchers to use them.
The Head of the Unit participated in UPF and CRG masters courses, as well as flow cytometry courses at other institutions in Barcelona. The CRG-BD Multicolour Flow Cytometry Workshop was hosted as a training workshop to advance in multicolour panel design for multiparametric flow cytometry assays. National collaborations also began in 2015 (with UPF, CRG and Universitat de Girona).