Flow Cytometry – GIMM Flow Cytometry – GIMM
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Flow Cytometry

Flow Cytometry

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The Flow Cytometry Facility provides a state of the art flow cytometry service and expertise to GIMM researchers as well as external groups from other research institutes and companies. 

Laboratory staff is well trained in instrument operation, providing support to cell sorting, sample acquisition and data analysis. Training is available for all analytical instruments and SOP are implemented to comply with the highest quality standards required to ensure reproducibility in science.

The need to find solutions to support research projects drives a continuous development of the facility, which aims to follow the advances in the flow cytometry field, collaborates with innovative projects, creates novel tools and methods to advance research, and implements strategies to improve the quality of the provided services.

Flow Cytometry info

  • Maintenance and Quality Control of scientific equipment – The Flow Cytometry Facility has integrated maintenance and quality assurance protocols for the operation of all scientific instruments.

 

  • Cell Analysis and Imaging Flow Cytometry – Available 7 days a week, 24 hours a day for use by trained researchers upon reservation on the online booking system.

 

  • Cell Sorting – Available for booking from 10am to 6pm from Monday to Friday.  The cell purification service is provided by a dedicated technician upon reservation on the online booking system.

 

  • Training in Flow Cytometry – The Flow Cytometry Facility provides lectures and hands-on sessions to all users, offering high quality training in flow cytometry, including polychromatic, spectral and imaging flow cytometry, as well cell sorting and data analysis.

 

  • Data Analysis – The facility provides a data analysis service, that includes training (FlowJo and FCS Express), consultation and optimized workflows to analyze flow data more efficiently. The facility also provides a limited number of one-year licenses for FlowJo and access to workstations with different data analysis software (FCS Express, FlowJo, IDEAS, ModFitLT and SpectroFlo).

 

  • Flow Cytometry Experimental Consulting – The facility provides guidance and cutting-edge inputs on projects, experimental planning, panel design, sample preparation, data analysis and data presentation regarding Flow Cytometry experiments.

Cell Sorters

  • BD FACSAria IIu – Oeiras – The BD FACSAria IIu is equipped with three lasers (405 nm, 488 nm, 640 nm), ten fluorescence detectors (2V, 6B, 2R) and two scatter detectors. It allows cell sorting of up to four populations simultaneously to eppendorfs, 5mL tubes or PCR tubes; two populations simultaneously for 15mL falcons; or one population to PCR strips, 6, 12, 24, 48, 96, 384-wells plates, to terazaki plates or to microscope slides. It has 70 μm, 85 μm and 100 nozzles available and can be used to sort BSL-2 samples.
  • BD FACSAria III – Lisbon – The BD FACSAria III is equipped with four lasers (405 nm, 488 nm, 561nm, 633 nm), 16 fluorescence detectors (6V, 2B, 5YG, 3R) and two scatter detectors. It allows cell sorting of up to four populations simultaneously to eppendorfs, 5mL tubes or PCR tubes; two populations simultaneously for 15mL falcons; or one population to PCR strips, 6, 12, 24, 48, 96, 384-wells plates, to terazaki plates or to microscope slides. It has 70 μm, 85 μm, 100 μm and 130 μm nozzles available and can only be used to sort BSL-1 samples.
  • BD FACSAria Fusion – Lisbon – The BD FACSAria Fusion is equipped with four lasers (405 nm, 488 nm, 561nm, 640 nm), 16 fluorescence detectors (6V, 2B, 5YG, 3R) and two scatter detectors. It allows cell sorting of up to four populations simultaneously to eppendorfs, 5mL tubes or PCR tubes; two populations simultaneously for 15mL falcons; or one population to PCR strips, 6, 12, 24, 48, 96, 384-wells plates, to terazaki plates or to microscope slides. It has 70 μm, 85 μm, 100 μm and 130 μm nozzles available and can be used to sort BSL-2 samples.
  • BD FACSymphony S6 SE – Lisbon – The BD FACSymphony S6 SE is a high parameter cell sorter equipped with five lasers (349 nm, 405 nm, 488 nm, 561 nm and 633 nm), 48 fluorescence and two scatter detectors. It allows cell sorting of up to six populations simultaneously to eppendorfs, 5mL tubes or PCR tubes; four populations simultaneously for a combination of 5 mL tubes and 15 mL falcons; or one population to PCR strips, 6, 12, 24, 48, 96, 384-wells plates, to terazaki plates or to microscope slides. It has 70 μm, 85 μm, 100 μm and 130 μm nozzles available and can be used to sort BSL-2 samples. The BD FACSymphony™ S6 SE allows for high-parameter spectral flow cytometry. The spectral enabled technology allows unmixing and compensation workflows. It enables you to gain full visible spectrum coverage to collect all light emitted without the need for filter changes, increasing dye flexibility and number of simultaneous colors. It also allows for autofluorescence unmixing, that may enable the resolution of difficult cells and populations.
  • Cytek Aurora CS – Oeiras – The Cytek Aurora CS is a full spectrum flow cytometer cell sorter equipped with five lasers (355 nm, 405 nm, 488 nm, 561 nm, 640 nm), 64 fluorescence and three scatter detectors. It allows cell sorting of up to six populations simultaneously to eppendorfs, 5mL tubes or PCR tubes; two populations simultaneously for 15 mL falcons; or one population to PCR strips, 6, 12, 24, 48, 96-wells plates, to microscope slides. It has 70 μm and 100 μm nozzles available and can be used to sort BSL-2 samples. The Aurora system provide flexibility, enabling the use of a wide array of fluorochrome combinations without reconfiguring the system for each application. It delivers high-resolution data at the single-cell level to resolve the most challenging cell populations, such as cells with high autofluorescence or low levels of expression of key biomarkers, regardless of assay complexity.

Cell Analyzers

  • BD Accuri C6 Plus – Lisbon – The BD Accuri C6 Plus cell analyzer is an easy-to-use benchtop flow cytometer that allows for absolute cell counting and has a flexible optical layout. It is equipped with two lasers (488 nm, 640 nm), two scatter detectors and four fluorescence detectors (3B, 1R).
  • BD LSRFortessa SORP – Oeiras – The BD LSR Fortessa SORP is equipped with three lasers (488 nm, 442 nm, 561 nm), one FSC-PMT for small particle analysis, two SSC detectors and ten fluorescence detectors (3B, 2BV, 5YG). It is coupled to a High Throughput Sampler (HTS) for sample acquisition in 96-well plates.
  • BD LSRFortessa – Lisbon – The BD LSR Fortessa is equipped with four lasers (405 nm, 488 nm, 561 nm, 640 nm), two scatter detectors and thirteen fluorescence detectors (4V, 2B, 5YG, 2R). It is equipped with a High Throughput Sampler (HTS) for sample acquisition in 96-well plates.
  • BD LSRFortessa X-20 – Lisbon – The BD Fortessa X-20 is equipped with four lasers (405 nm, 488 nm, 561 nm 640 nm), two scatter detectors and 16 fluorescence detectors (6V, 2B, 5YG, 3R).
  • BD LSRFortessa X-20 – Oeiras – The BD Fortessa X-20 is equipped with four lasers (405 nm, 488 nm, 561 nm, 640 nm), two detectors and 16 fluorescence detectors (6V, 2B, 5YG, 3R).
  • BD FACSymphony A5 SE – Lisbon – The BD FACSymphony A5 SE is a high parameter cell analyzer equipped with five lasers (349 nm, 405 nm, 488 nm, 561 nm and 640 nm), 48 fluorescence detectors and two scatter detectors. It allows for high-parameter spectral flow cytometry.  The spectral enabled technology allows unmixing and compensation workflows. It enables you to gain full visible spectrum coverage to collect all light emitted without the need for filter changes, increasing dye flexibility and number of simultaneous colors. It also allows for autofluorescence unmixing, that may enable the resolution of difficult cells and populations.
  • Cytek Aurora – Oeiras – Cytek Aurora is a full spectrum flow cytometer equipped with four lasers (405 nm, 488 nm, 561 nm, 640 nm), three scatter detectors and 48 fluorescence detectors. It has installed an Auto Micro-Sampling System (AMS) for sample acquisition in 96-well plates.  The Aurora system provide flexibility, enabling the use of a wide array of fluorochrome combinations without reconfiguring the system for each application. It delivers high-resolution data at the single-cell level to resolve the most challenging cell populations, such as cells with high autofluorescence or low levels of expression of key biomarkers, regardless of assay complexity.
  • Cytek Aurora – Lisbon – Cytek Aurora is a full spectrum flow cytometer equipped with five lasers (355 nm, 405nm, 488 nm, 561 nm, 640 nm), three scatter detectors and 64 fluorescence detectors. It has installed an Automated Sample Loader (ASL) with an orbital shaker and compatible with 96-well plates , 96-deep well plates and 40-tube racks.  The Aurora system provide flexibility, enabling the use of a wide array of fluorochrome combinations without reconfiguring the system for each application. It delivers high-resolution data at the single-cell level to resolve the most challenging cell populations, such as cells with high autofluorescence or low levels of expression of key biomarkers, regardless of assay complexity.

 

Imaging Flow Cytometer

  • Amnis ImageStreamX – Lisbon – Benchtop multispectral imaging flow cytometer equipped with two CCD cameras that can acquire up to 12 images simultaneously of each cell or object in brightfield, scatter and multiple fluorescent channels.

Cell Counters

  • Scepter Millipore – Oeiras – The Scepter is a portable high precision cell counter that provides true automation that accompanies vision-based systems. It uses disposable sensors that can be provided by the facility.

Data Analysis Resources

  • MacFlow 2 – Lisbon – Data analysis workstation with multiple software installed, including: FlowJo and FCS Express.
  • Moonwalk Data Analysis Workstation – Oeiras – High-performance data analysis workstation dedicated to analysis of spectral flow cytometry data with SpectroFlo.
  • PC Workstation – Lisbon – Data analysis workstation with multiple software installed, including: FACSDiva and ModFitFL.
  • Tango Data Analysis Workstation – Oeiras – Remote high-performance data analysis workstation, with multiple software installed, including: FlowJo Milliplex Analyst, IDEAS, among others.

The Flow Cytometry Facility systems are available to GIMM researchers and external users.

All users must be registered in Agendo, our online booking system.

Please contact us (mariana.fernandes@medicina.ulisboa.pt) for more information.

If you used our services, please acknowledge us in publications or other formal presentations. This is one of our key performance indicators and very important for future funding. You are welcome to use the following examples:

In Materials and Methods: “All experiments were performed at the Flow Cytometry Facility of Gulbenkian Institute for Molecular Medicine.”

In Acknowledgements: “We would like to thank the Flow Cytometry Facility of Gulbenkian Institute for Molecular Medicine for their technical support.”