The Flow Cytometry and Cell Sorting Unit offers technical support and guidance to ISPA’s research groups and visiting researchers who are interested in flow cytometry for experimental design, selection of markers, sample acquisition and analysis, and discussion of the results obtained.
The main services we offer are:
- Phenotypical studies through marking surface and intracytoplasmic antigens.
- Measuring cell viability through the use of viability markers (PI, 7AAD, fixable viability dyes, etc).
- Identifying cell apoptosis in response to bioactive compounds (Annexin V, Caspase 3).
- Identifying genetically-modified populations through reporter genes (mainly GFP).
- Identifying the various phases of the cell cycle using propidium iodide (PI).
- Quantifying cell proliferation (BrdU, Ki-67, etc)
- Measuring autophagy in cell lines (measurement of the LC3 protein, CytoID©, etc.)
- Soluble molecule studies using microspheres (cytometric bead array “CBA”)
- Identification of microvesicles using magnetic microspheres.
- Isolation of cell populations (in tubes/on plates/ single cell) using FACS (Fluorescence Activated Cell Sorting) technology.
The Unit’s work also includes the development of new technology and evaluation of new equipment, organisation of training activities and educating users about flow cytometry.
Laboratorio Citometría N-1 F.40. Sede ISPA
Av. Hospital Universitario s/n 33011-Oviedo, Spain.
Telephone: +34 985652413 (Extension 39793)
Cristina Martín Martín
- Doctor of Biosciences. Specialist Health Professional (FEA) in Immunology
- Technician specialised in flow cytometry and sorting
- Member of the Sociedad Ibérica de Citometría (Iberian Cytometry Society)
ISPA’s Flow Cytometry Unit contains a FACSAria IIu flow cytometer, which is suitable for fluorometric studies of up to eight parameters and electrostatic cell sorting. It consists of:
- A 488nm laser (with FITC, Pe, PerCP/Pe-Cy5/PercP-Cy5.5, Pe-Cy7, forward scatter [FSC] and side scatter [SSC] detectors), also compatible with Alexa 488, PI, 7AAD, CFSE, GFP and Rhodamine 123.
- A 633 nm laser (with APC and APC-H7/APC-Cy7 detectors), compatible with DRAQ5, DRAQ7, Alexa 647 and Alexa 750.
- Sort block with capacity for four simultaneous collection tubes and isolation on a plate (including single-cell capability).
- Facs Diva V8.02 analysis software, which incorporates the CST quality control module to optimise voltage and monitor the operational parameters of the device.
- FlowJo V10 analysis software, suitable for analysis of all cytometry files with the .fcs extension. Able to export statistics, modify compensations, compress ACS (Archival Cytometry Standard) files, and perform other functions.
- Users may request use of the cytometer for cell acquisition or sorting, or of the analysis platform alone, by consulting a member of the Unit’s staff from Monday to Friday between 9am and 4pm.
- The device is to be used and handled only by service technicians.
- If you need to use the device outside these hours, it is possible to make an appointment to do so.
- Before requesting use of the Cytometry Platform’s services, we recommend consulting the Recommendations Before Making a Request document.
- An application form must be completed for each request, which should be signed by the principal investigator and submitted to a member of staff in the Unit (either in person or by e-mail): Cytometry service request.
- If you are using the service for the first time, you will also need to fill in the Biosafety Questionnaire.
- The date and time assigned will depend on availability and compatibility with other reservations. Reservations will be made by the Unit’s staff, on a strictly first-come first-served basis.
- Only reservations made at least three working days in advance will be accepted.
- Given that all types of reservations require the device to be started up and calibrated, cancellations must also be made with one day’s notice. If this condition is not met, a charge will be made, according to the established fees.
|SERVICE||ISPA||Other public organisations||Standard fee|
|Analysis of .fcs files||€3/hour||€5/hour||€10/hour|
|Acquisition of prepared samples||€15/hour||€25/hour||€ 35/hour|
|Sorting of prepared samples||€45/hour||€75/hour||€100/hour|
(Prices do not include VAT)