Flow Cytometry and Cell Sorting

Flow Cytometry and Cell Sorting

  1. Introduction
  2. Techniques
  3. Equipment
  4. Usage rules
  5. Prices
Flow Cytometry and Cell Sorting

The Flow Cytometry and Cell Sorting Facility is a specialized unit in cell analysis techniques that aims to provide IBiS researchers with the latest technology in flow cytometry, which will serve as a support to their biomedical research.

Location:1st Floor

Working hours:M-F: From 9 a.m. to 5 p.m. *

(*) Those experiments that need time past the established hours could be made prior knowledge of the technician in charge.

Cell analysis applications provided by this service are numerous and diverse. The most common application in flow cytometry is to analyze the different cell phenotypes by distinguishing the differential expression of cell surface proteins. In addition to this, a number of different parameters and processes can be analysed also by flow cytometry, and hence can be performed in our Service.

Among the possible assays available we highlight a few:

  • Cellular immunophenotyping.
  • Analysis of cell cycle, proliferation and apoptosis.
  • Separation of cell populations by flow cytometry (cell sorting).
  • Multiplexed Analysis.
  • Analysis of microparticles.
  • Side Population Analysis.
  • Analysis of ROS.
  • Kinetics.
  • Development of novel flow cytometric techniques for the measurement of different parameters, including nuclear, cytoplasmic and extracellular parameters.
Click on the different equipment for details on features.

Canto II (Becton Dickinson) analyzer.

 

  • Lasers:
    • Violet: 405 nm 30mW. Two detectors.
    • Blue: 488nm 20mW. Five detectors (plus FSC).
    • Red: 640 nm 17 mW.
  • High Throughput Sampler (96w, 384w).

 


TO CHECK THE OPTIC CONFIGURATION, PLEASE, SEE THE NORMATIVE.

LRS II Fortessa cytometer analyzer (Becton Dickinson)

 

  • Lasers:
    • Violet: 405 nm 50mW. Seven detectors.
    • Blue: 488nm 50mW. Three detectors (plus FSC).
    • Red: 640 nm 40 mW. Three detectors.
    • Green: 561nm 50mW. Five detectors.
  • High Throughput Sampler (96w, 384w).

 

 
TO CHECK THE OPTIC CONFIGURATION, PLEASE, SEE THE NORMATIVE.

BD FacsJazz (Becton Dickinson) Cell Sorter

 

  • Jet-in-air with 100 um nozzle.
  • Lasers:
    • Blue Laser (488nm, 80mW).  Three detectors (plus FSC).
    • Green Laser (561nm, 50mW). Four detectors.

 

TO CHECK THE OPTIC CONFIGURATION, PLEASE, SEE THE NORMATIVE.
 

Facs Aria Fusion (Becton Dickinson) Cell Sorter

 

  • Lasers:
    • UV: 355nm, 15mW. Two detectors.
    • Violet: 405nm, 85mW. Six detectors.
    • Blue: 488nm, 50mW. Three detectors (plus FSC).
    • Red: 633 nm; 100 mW. Three detectors.
    • Green: 561 nm, 50mW. Five detectors.
  • Several racks for adquisition and collectection tubes.
  • Control temperature system for adquisition and collection samples.
  • Four ways separation
  • Class II Biosafety Cabinet to sort in aseptic conditions.
  • Index sorting system, to find sorted cells in the analysis plot, improving next separations (available only in plate separations).

 

TO CHECK THE OPTIC CONFIGURATION, PLEASE, SEE THE NORMATIVE.
 

Multi-Range Large Particle Flow Cytometer (Union Biometrica)

The BioSorter® is a continuous flow system capable of analyzing, sorting and dispensing objects ranging in size from 10 to 1,500 µm, depending of the Fluidic and Optics Core Assemblies (FOCA).

Available FOCA: 1,000 µm

Multi-Range Large Particle Flow Cytometer (Union Biometrica)

Magnetic Automatic Separator AutoMACS ProSeparator (Miltenyi Biotec)

 

  • Two magnetic columms.
  • Different racks for sample adquisition.

As general rule.

In order to make online reservations in the Flow Cytometry and Cell Separation Facility of IBiS, it is necessary for the user to be previously signed up in the online booking system https://servicios.ibis-sevilla.es/

Equipment located in the first room is user-friendly and those in the second room are necessarily assisted by the facility technician.

First-time training is mandatory for those instruments that can be used by the researchers.

There are some basic hygienic standards that must be respected:

  • Access to the service is restricted, especially for the cell separator room.
  • It is strictly forbidden to eat, drink, or place food in the flow cytometry room.
  • The use of gloves is required.
  • When using the centrifuge with biohazardous samples, capped tubes should be used.

FREE ACCESS EQUIPMENT: FORTESSA, CANTO II, AUTOMACS.

  • These instruments will be handled by the researchers themselves, who must know the usage of them. The facility technician will provide appropriate training to researchers who apply for the first time to the use of these equipments, and researchers will not be able to use these equipments until they have the OK from the technician, which will be credited through THE UNLOCK of the automatic reservation with the technician in the reservation platform.
  • The setting-up of the devices at the beginning of each day (including calibrations and washings) will be performed by the facility technician, making them available for use.
  • It is the obligation of the researcher to leave the equipment clean and ready for the next user. The closing and cleaning of the unit is the responsibility of the LAST REGISTERED USER ON THE CALENDAR, whether it has been the last one using the device or not.
  • The user who arrives late at the reservation, can start the experiment as long as it does not delay the next user.
  • A maximum of 15 min will be granted to researchers who could not complete the experiment before the next reservation. If it takes longer, they automatically have to give way to the next researcher, but they may finish the experiment at the end of the day if there are free slots, or they will have to make a new reservation for the next day.
  • Changes and cancellations in reservations for the same day will not be accepted when they involve an unnecessary expense to the service.
  • The analysis softwares are installed on the computers of the users (to request the installation contact the computer technician). They all need a license to run the program. There are 8 licenses for the DIVA v8.0 software and 4 for the FlowJo v10.2 software. These licenses will be requested from the service technician and the loan will be for one week.

HANDLED BY SERVICE TECHNICIAN EQUIPMENT: CKX41 NVERTED FLUORESCENCE MICROSCOPE, CELL SORTERS FACS ARIA FUSION, FACS JAZZ & UNION BIOMETRICA BIOSORTER.

  • Cell sorters will be handled solely by the facility technician, so their use has to fit with the technician schedule.
  • The use of the inverted fluorescence microscope is restricted to users of the cell sorter.
  • The maintenance and cleaning of the cell sorter is responsibility of the technician.
  • Booking this equipment will be considered as a technician service, so it will have to be validated once the operator agrees. The inobservance of the schedule for this instrument will lead to the cancellation of the experiment.
  • Keep in mind that the equipment configuration and optical characteristics of each experiment can be different (size of the nozzle, outlet pressure, flow rate, calculation of the drop delay, etc.), so it is ideal to free a period of 30 minutes between one reservation and the next. This waiting time is not always necessary, so users are not required to comply, except in cases in which the technician asks for it.
  • For specific and useful information before separating cells, please consult the facility technician.

Administrative Regulations.

  • The billing will be applied based on the latest published rates, approved by the IBiS Management.
  • The billing will be applied quarterly, unless otherwise specified by the IBiS Management.
  • A copy of the bill will be mailed to each IR in those groups who use the flow cytometry facility. This email must be returned to the facility technician after proper agreement, or highlighting any possible disagreements.
  • The effectiveness of cytometry softwares in the equipments gets worse with data accumulation. Therefore, each time the users make use of an instrument of flow cytometry, they should export the data to the server as indicated by IBiS computer facility.
  • However, the flow cytometry technician will regularly and systematically remove the data stored in the equipments, once or twice a year depending on the saturation of the equipment.
  • The removal of data will be informed in advance and repeatedly via email.

For information about fees and charges applicable to our services, please contact: majocastro@us.es.