Cytometry
Plataforma de Citometria
Planta Baixa IGTP Edifici Mar
Institut de Recerca Germans Trias i Pujol (IGTP)
Crta. de Can Ruti – camí de les escoles, s/n
08916 Badalona, Barcelona, Spain
Services
Cell separation
Separator flow cytometers (FACSAria II) allow cells to be separated according to parameters previously defined by the operator. To purify different cell populations, flow cytometry is specially designed for applications that require a high purity of collection. The use of multiple fluorochromes simultaneously allows to classify and separate cell subpopulations based on the expression of different cellular properties.
The preparation of the cell separation experiment requires a series of steps prior to its performance that must be specified in advance with the head of the Cytometry Platform.
Acquisition and analysis with flow analyzer cytometer
BD FACSCanto II, BD FACSLyric and BD LSR Fortessa are the analyzing cytometers that the user can use to carry out different studies. These equipments can be used directly by the user, either accrediting experience in the handling of these equipments or by means of a previous course on the part of the personnel of the platform of Cytometry. Additionally, the FACSAria II cell separator can be used as an analyzer cytometer, especially in cases of the need to use more fluorescence detectors.
Offline analysis in Mac / Windows environment with FACSDiva and FlowJo software, offline compensation and “Logicle” or toponential scales and ModFit.
Training
Design of customized courses. Several courses on the fundamentals and applications of flow cytometry are held annually at the IGTP.
Consulting
The Cytometry platform provides users with a consulting service on issues related to flow cytometry.
Equipment
Cell Separation
- FACSAria II Cell Separator (BD Biosciences)Three lasers (405, 488 and 633 nm), fifteen detectors (thirteen fluorescences and two light scattering parameters). ACDU (Automated Cell Deposition Unit) for separation into different types of well plates and slides; single cell option. Installed options: aerosol evacuation system (AMO) and temperature control for refrigerated bath. FACSDiva software.
- Flow cytometers analyzers
LSRFortessa SORP Flow Cytometer (BD Biosciences)Equipped with blue solid state laser (488nm), red He-Ne laser (640 nm), violet laser (405 nm) and green laser (532 nm), it allows to work with 18 fluorescences and 2 dispersion parameters (FSC and SSC ), analyzing for each the area, height and width of the electric pulse. Digital signal processing, acquisition and analysis with FACSDiva software. HTS system for the acquisition of samples with 96-well plates. - FACSLyric Cell Separator (BD Biosciences)Equipped with solid state blue laser (488nm), red He-Ne laser (633 nm) and violet laser (405 nm), it allows to work with ten fluorescences and two dispersion parameters (FSC and SSC). Digital signal processing system with BD FACSSuite software. Equipped with automated sample loader for 96-well tubes and plates.
- FACSCanto II Flow Cytometer (BD Biosciences)Equipped with blue solid state laser (488nm), red He-Ne laser (633nm) and violet laser (405nm), it allows to work with eight fluorescences and two dispersion parameters (FSC and SSC). Digital signal processing system with FACSDiva software. Equipped with automated sample charger for tubes (Loader).
- Luminex® 200 (Luminex Corp.)The Luminex® 200 system is an analyzer based on the principles of flow cytometry. Luminex® 200 sets the standard for multiplexing and offers the ability to perform up to 100 different tests in a single reaction volume and use magnetic or polystyrene beads. It gets great sample performance with a plate reading time of 96 jars of just 45 minutes.
Programes per anàlisi off-line
Software
FlowJo, FACSDiva and CellQuestPro installed on an iMac 27 “Intel i7 16 GB RAM computer with MAC OSX and Windows systems (XP and 7).
High dimensional spectral cytometry
Aurora 5-laser (Cytek Biosciences)
Installed in May 2020, it is configured with five lasers (355, 405, 488, 561 and 640 nm), three scattering parameters and 64 fluorescence channels. The Aurora allows high-dimensional data with designed panels (40 colors simultaneously and beyond). Equipped with an automatic micro-sampling system (AMS) for the acquisition of high performance with 96-well plates. Data analysis with SpectroFlo software to differentiate spectral raw data.
Publications
Methods Mol Biol. 2022;2386:43-60.
