Flow Cytometry Core Facility

The BD FACSCalibur is a six parameter instrument equipped with two lasers, a 488-nm argon-ion laser and a 635-nm red diode laser, capable of detecting 4 fluorescent signals.

The BD FACSAria is a sixteen parameter instrument equipped with four fiber-optic-linked solid-state lasers, a 488-nm blue Sapphire laser, a 633-nm red HeNe laser, a 405-nm violet laser, and a 335-nm UV laser, capable of detecting 14 fluorescent signals. The FACSAria can  two- and four- way bulk sorting for a variety of tube sizes as well as automatic cell deposition (ACDU) for sorting into multiwell plates or onto microscope slides. The FACSAria is also equipped with aerosol management (AM) and Temperature Control (TC) options.

Flow cytometry has two general applications; quantitative analysis and cell sorting.

Analysis and differentiation of the cells is based on physical characteristics such as cell size and granularity (internal complexity) and any cell component or function that can be detected by a fluorescent compound either in the form of antibodies or dyes.

Cell sorting involves physical separation of distinct subpopulations of interest on the basis of the measured characteristics for the purpose of performing further experimental procedures.

Perhaps the most common use is the identification of the presence of antigens either on the surface of or within cells. However, flow cytometry may be used for a number of studies on cells including:

• Multicolor analysis
• Apoptosis
• Rare Event Analysis
• Sorting
• FRET
• Cycle Analysis
• Functional Analysis
• Translocation Assay
• Single Cell Cloning

Location and Hours of Operation: the Flow Cytometry Core Facility is located in the Charles E. Schmidt College of Medicine at 777 Glades Road, Boca Raton, building 71 in room 320 on the third floor. The facility is open for data acquisition/analysis and cell sorting Monday through Friday from  9 a.m. to 6 p.m.

Scheduling Procedures: users must complete/submit an instrument request form to the Flow Cytometry Core Facility for the FACSAria and FACSCalibur cytometers prior to the requested appointment date (see links below). Users will be contacted in advance to ensure that they can be accommodated. Instrument request example forms for FACSAria and FACSCalibur are available below.

FACSAria instrument request form

FACSCalibur instrument request form

Instrument request example forms

Cancelation Procedures: cancelations are allowed up to 24 hours prior to the scheduled time. Failure to cancel prior to 24 hours will result in a charge for a half-hour at the appropriated billing rate. No-shows will be charged for the full amount of the time reserved.

• A daily FACS Comp and CS&T; file will be recorded on the FACSCalibur and FACSAria to ensure optimal performance.
• Facility personnel run the samples and provide the users with reports of data analysis. Upon request, facility personnel will provide help for generation of publication-quality graphics.
• All experimental files generated on the FACSCalibur and FACSAria will be archived according to the standard file naming structure established by the facility.
• The FACS Director is available Friday afternoons to discuss with users their experimental design, controls, and other needs to help meet their goals. The Director can also provide input into analysis, data interpretation, and advice for future directions.

The Flow Cytometry Core Facility operates on a Fee-for-Service basis, which includes maintenance, data acquisition, analysis, and consultation.

 ( Rates are fixed by the FACS Steering Committee)

• There is no charge for the time needed to sterilize the machines or to set up the 96-well single-cell acquisition.
• Users are not billed for the time needed for instrument repair or configuration unless this results from poor user sample quality (i.e. sticky or clumpy cells) or a failure of the user to adequately communicate required analysis parameters in advance.
• At the end of each month, the Core Director will submit a list of service charges to Research Finance sorted by Principal Investigator and the monthly charge amount. A monthly statement will be sent to each PI. Research Finance will debit the corresponding account number and credit all revenue to the Flow Cytometry Core Facility budget.