COBRE Center for Stem Cells and Aging
Flow Cytometry Core
The Rhode Island Hospital Stem Cells and Aging (SCA) COBRE Flow Cytometry Core function is to provide high quality, cost effective, state-of-the art flow cytometry and multiparameter cell sorting instrumentation and associated expertise and services to all investigators in the SCA COBRE and to the general research community in Rhode Island.
COBRE Flow Cytometry Core Staff and Services
The core is staffed by Mark Dooner and Loren Fast, PhD
Mark Dooner directs and manages the core and serves as operator for the BD Biosciences Influx cell sorter and LSRII. Dr. Fast serves as co-director and is available as an advisor to the investigators in experimental design and data interpretation. Together they have over 45 years of flow cytometry experience.
The core has had experience with a variety of different flow cytometry applications and protocols and will work with both experienced and novice investigators to help plan and execute successful flow cytometry experiments.
The Rhode Island Flow Cytometry Facility is in the Coro West Building, 5th Floor, Suite 5.08A, One Hoppin Street, Providence.
|Instrument or Service||Price|
|LSRII assisted or outside users||$65/hour|
|Influx cell sorting||$75/hour|
|Training (LSRII) 1 hour||$50|
Facility Oversight and Contact Information
All first-time users must contact Mark Dooner to discuss their project and to set up a user account. Every effort will be made to accommodate sorting requests in a timely manner. Booking in advance is recommended.
Please note SCA COBRE projects and pilot projects will be given priority.
Mark Dooner, COBRE Flow Cytometry Core, Rhode Island Hospital
Flow Cytometry Core Equipment
BD Biosciences Influx Cell Sorter
- 5 laser system (355nm, 405nm, 488nm, 561nm and 638nm) for simultaneous detection of up to 14 parameters.
- Sorts up to 4 different cell populations.
- Capable of single cell deposition into many different sized culture plates (96 or 384 well plates, etc).
- Equipped with a small particle objective for FSC size determination down to 0.5 µm.
- Enclosed in a biosafety hood.
The sorter separates particles on the detection of fluorescent markers on particles such as cells, chromosomes, bacteria or nuclei.
Then the collected cells are suitable for downstream analysis either in molecular applications like protein assays and PCR, or used in viable assays such as tissue culture or injected into in vivo models.
Generated data may also be used to determine percentages of interrogated populations. The sorter can separate populations into tubes (1.5ml, 4ml, 15ml, 50ml), multi-well plates (6, 24, 48, 96, 384), or directly onto microscope slides.
Nanosight Nanoparticle Counter
- Examines size, size distribution and concentration.
- Fluorescent adapter for extracellular vesicles (EVs) surface receptor studies.
- Able to control for microvesicle number in experiments, and possibly identify subpopulations of EVs based on surface receptors
- Particles are illuminated using a laser through a prism, and light scatter is captured on a CCD camera.
- Nanoparticle tracking analysis (NTA) – direct and real-time visualization and analysis of nanoparticles in liquids, and calculate the size, size distribution and concentration of the particles. Brownian motion of nanoparticles is analyzed using the Stokes-Einstein equation.
BD Biosciences LSRII Analyzer/Flow Cytometer
- Digital, 4 laser (355nm, 405nm, 488nm and 635nm) flow cytometric analysis system with 10 fluorescent channels plus 2 scatter channels.
- BD FACSDiva software with digital compensation and ModFit software for cell cycle analysis.