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ICMB

Cell Sorting Flow Cytometry

Instruments:
BD FACSAria Cell Sorter
The FACSAria flow cytometer is used for cell sorting only. For non-sort applications the FACSCalibur is available.

 

The FACSAria is a benchtop high-speed sorter equipped with three lasers providing excitation sources of 375-nm, 488-nm, and 633-nm wavelengths. Digital acquisition rates of up to 70,000 events/second. Multicolor analysis of up to 11 parameters (Forward Scatter, Side Scatter, and 9 Fluorescence). Two- and four-way bulk sorting devices for microtubes, 12 x 75mm and 15mL tubes. Automated Cell Deposition Unit for sorting into multiwell plates (6,24,48,96, and 384) or microscope slides.

Most dyes designed for the FACSAria are specially manufactured by BD.We recommend you used the following fluorescent labels (only one dye can be used in each category):

UV Laser:

375 PMT A: (515-545 nm)
Alexa Fluor 430 - Excitation at 375 nm, emission at 541 nm.

375 PMT B: (430-470 nm)
DAPI - Excitation at 375 nm, emission at 461 nm.

Blue Laser:
488 PMT A: (750-810 nm)
PE-CY7 (phycoerythrin-Cy7) - Excitation at 488 nm, emission at 767 nm.
488 PMT B: (675-715 nm)
PI (Propidium Iodide) - Excitation at 488 nm, emission at 617 nm
PE-Cy5.5 (phycoerythrin-Cy5.5) - Excitation at 488 nm, emission at 667 nm.
PerCP (peridinin chlorophyll protein) - Excitation at 488 nm, emission at 678 nm.
PerCP-Cy5.5 (peridinin chlorophyll protein) - Excitation at 488 nm, emission at 695 nm.
488 PMT C: (600-620 nm)
PE-TXRed (phycoerythrin-TexasRed) - Excitation at 488 nm, emission at 578 nm.
488 PMT D : (564-606 nm)
PE (phycoerythrin) - Excitation at 488 nm, emission at 578 nm.
488 PMT E:
(515-545 nm)
FITC (Fluorescein) - Excitation at 488 nm, emission at 519 nm.
Alexa Fluor 488 - Excitation at 488 nm, emission at 519 nm.
GFP (Green Fluorescent Protein) - Excitation at 488 nm, emission at 508 nm.

Red Laser:
633 PMT A : (750-810 nm)
APC-Cy7 (allophycocyanin-Cy7) - Excitation at 633 nm, emission at 785 nm.
633 PMT B : (650-670 nm)
APC (allophycocyanin) - Excitation at 633 nm, emission at 660 nm.
AlexaFluor 647- Excitation at 633 nm, emission at 668 nm.

Access and Training:

Training is not available for the FACSAria. This instrument is only available via "Assisted Use".

For more information about sorting, stop by the Microscopy Facility Office (1.426H) and pick up a Sort Request form.


Location:
The FACSAria is located in NMS 2.114.
Core Facilities Map

Instrument Supervisor:
Please contact Richard Salinas for information.

Fees:

ICMB charges all campus users $55.00/ hour for use of the FACSAria.


Flow Cytometry Information and Links:

BD Biosciences:
FACSAria Information
BD-Pharmingen-Protocols
BD-Fluorchrome Information


General Cytometry Information
Purdue University Cytomery Labs (PUCL)
PUCL Lecture Slides

Clinical Cytometry Scoiety
ISAC Cytometry Page
Protocols Online
Flow Cytometry at UT


Cell Cycle Information

Cell Cycle Fundamentals
- Written by Nate Regimbal Biogen Idec, Inc.


Cytometry Freeware

WinMDI Version 2.8
A very good, easy to use, software for general analysis. This freeware is provided by Joseph Trotter of the Scripps Research Institute, La Jolla, CA. Check out Purdue's WinMDI tutorial.

Cyflogic
Another easy to use, general analysis program that is "windows-only".

Flowing Software
A new, general analysis program that is also "windows-only".