Flow cytometry is a powerful technology used for characterizing and analyzing cells. The fluidics, optics, and electronics systems in flow cytometry work together to simultaneously measure and analyze multiple physical characteristics of particles, usually cells, as they move in a fluid stream through a beam of light. A flow cytometer can tell us about a particle’s relative size, fluorescence intensity, and granularity or internal complexity. The BC Children's Hospital Research Institute Flow Core Facility provides equipment for flow cytometric analysis and full service cell sorting, as well as flow cytometry training and education for new users.
The ability of flow cytometers to evaluate particles or cells at an extremely rapid rate (e.g. up to 25,000 events per second) makes this technology ideal for quantitatively analyzing the physical properties of target cells and particles. Any 0.2-50 micrometer particles or cell suspension of peripheral blood, bone marrow, body fluid or tissue can be labelled with multiple colour antibody cocktails and run through the multi-laser flow cytometer for FACS analysis.
FACS has been widely used in applications such as: tracking GFP (YFP, CFP) expression, subset identification, cell activation status, cell cycling, surface protein expression, cytokine production, chemokine receptor repertoire, cell to cell interactions, cell signaling, cell viability and apoptosis.
The Research Institute Flow Core Facility has the following analyzers:
This high performance bench top flow cytometer system by Beckman Coulter was purchased in 2016 and installed with 3 lasers (405nm, 488nm, 640nm). It has the flexibility to allow 10 colour experiments and simple reconfiguration with filter swapping. It uses a 96 well HTS plate loader module accepts U, V or flat bottom plates and a semi-automatic single tube loader (1.5ml/5ml tubes) with an option for custom tubes. It can display data with 7 logs of dynamic range. The license free acquisition and analysis software (CytExpert) allows remote installation for offline set up and analysis.
- BD LSRFortessa X-20
This compact flow cytometry analyzer was a specially ordered research instrument. It was purchased in the middle of 2014 and was upgraded to feature 18 fluorescence detectors with a total of 20 parameters after the installation. It has 405nm, 488nm, 561nm and 640nm lasers. The instrument uses Diva 8 software under the manufacture's full service warranty.
- BD LSRII
The LSRII was also a specially ordered system in 2009. It has been equipped with 480nm, 635nm, 532nm and 405nm lasers, and is capable of measuring 18 fluorescence colours with a total of 20 parameters. This instrument uses FACS Diva 6 software under BD’s full service warranty.
- BD FACSymphony
The system is a novel cell analyzer which was a special order for us in Nov 2017. It installed 355nm UV, 405nm violet, 488nm Blue, 561nm yellow-green and 640nm red lasers which leverages the inherent benefits of flow cytometry and enables the simultaneous measurement of up to 29 different characteristics of a single cell. This high parameter flow cytometer is a powerful analytical tool that enables researchers to identify and analyze distinctive phenotypes in heterogeneous populations using Diva 8 software.
- Fluoro Elispot
This Elispot reader was purchased in 2011 and can detect highly sensitive, microplate-based assays for cytokine secreting cells. It is equipped with two-colour plates and regular ELISA spot readers using AID software. It was made by AID Autoimmun Diagnostika GMBH (Germany).
One of the properties of flow cytometers is the ability to sort particles or cells by electronic deflection into a separate collection of tubes, plates or slides. Because multiple fluorochromes can be assessed simultaneously by flow cytometry, it can easily separate rare events from complex mixtures of cells on the basis of multiple marker expressions. Thus, flow cytometry is especially well suited for cell purification requiring high purity. The Research Institute Flow Core Facility has a high-speed cell sorter, BD FACS Aria IIu, which is capable of both analysis and cell sorting. After sorting, purified cells can be, for example, cultured, cloned, or processed for protein expression and function analysis, for mRNA analysis using quantitative PCR, or for gene array techniques, among many other possibilities.
- Beckman Coulter Astrios
The Astrios was purchased in 2016. This high speed cell sorter in a bio-safety cabinet provides level 2 biohazard sample and personal protection. Utilizing 4 Lasers with 20 fluorescent channels its two independent FSC detectors provide enhanced forward scatter and simultaneous sorting of small and large particles (from 0.2 microns to 30 microns). The Astrios is capable of up to 6-way sorting with temperature control and is “Jet-in-air” based for the improved viability of cells. Sample can be prepared in 0.5, 1.5, 5/7, 15 and 50mL tubes and collection options include 2way 15ml, 2way 50ml, mixed media 15/50ml, 6-way (5ml/1.5ml), and micro-plate-slide.
- BD FACS Aria
This FACS Aria was originally purchased in 2005 and it was updated to Aria IIu in March 2014. The FACS Aria has 404nm, 488nm and 633nm lasers and has been equipped with 11 fluorescence detectors. The instrument uses Diva 6 software and has been under BD’s full warranty service for the past few years.
- A new cell sorter model will be installed in 2016.
In addition to analysis and cell sorting, the BCCHR Flow Core Facility offers comprehensive training on instruments (FACS Canto, LSRII, Fortessa X-20 and FACS Aria) and software (CellquestPro, FACS Diva, FlowJo and FCS Express, etc). We can also consult on experimental design, fluocrome combinations and maximizing the effectiveness and cost of your experiments.
We aim to provide world-class flow cytometry services to BCCHR students, staff, researchers and scientists. We also welcome outside users. For more information, please contact Dr. Lisa Xu.
For more information or inquiries, contact firstname.lastname@example.org
To book the equipment or submit a service order, create your user profile on our online booking system at https://bcch-cts.calpendo.com/ and submit your grant authorization here for billing: https://rc.bcchr.ca/redcap/surveys/?s=9YP78F9JTT