Flow Cytometry
Instruments:
Becton and Dickinson FACSCalibur
Flow Cytometer
This flow cytometer is used for primarily for cell analysis with simultaneous acquisition of forward scatter, side scatter, and fluorescence. This is a dual-laser, four color system with very basic sorting ability. Excitation lines include a 488 nm and 635 nm. Emission bands are centered on 530 nm, 585 nm, 661 nm, and 670 nm. Software used with the cytometer include CellQuest Pro, FACSComp, and ModFit LT.
Most dyes designed for the FACSCaliber are specially manufactured
by BD. We recommend you used the following fluorescent labels:
FITC (Fluorescein) or Alexa 488 - Excitation at 488 nm, emission at 530 nm.
PE (phycoerythrin) Excitation at 488 nm, emission at 585 nm.
PerCP (peridinin chlorophyll protein) Excitation at 488 nm, emission at 670
nm.
APC (allophycocyanin) Excitation at 635 nm, emission at 661 nm.
Other dyes can be used on this system, please contact us if you have special
needs.
For information about training, stop by the Microscopy Facility Office (1.426
H) to discuss your flow cytometry plans and set up a training session. Each trainee
must attend at least 2 hours of training with Angela Bardo. Once training is complete the trained user will be given unassisted
access to the instrument.
Important note: This instrument will be housed in a Biosafety Level II room. You may be required to take additional safety courses through UT EH&S before you are trained.
Location:
The FACSCalibur is located in MBB
1.426.
Core Facilities Map
Instrument Supervisor:
Please contact Angela
Bardo for information.
Secondary contacts for this Instrument are
Cecil Harkey and Michelle
Gadush.
Fees:
ICMB charges all users $18.00/ hour
for use of the FACSCaliber.
Flow Cytometry Information and Links:
Becton and Dickinson:
FACSCaliber
Information
BD-Pharmingen-Protocols
General Cytometry Information
Purdue
University Cytomery Labs (PUCL)
PUCL Lecture
Slides
cytomics.info
Clinical Cytometry Scoiety
ISAC Cytometry Page
Protocol
Online
Flow
Cytometry at UT
Cell Cycle Information
Cell Cycle Fundamentals -Written by Nate Regimbal Biogen Idec, Inc. Posted on Purdue Site
Cytometry Freeware
WinMDI Version 2.8
This is 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.
Cylchred
This is an automatic cell cycle analysis program provided by UWCM in Cardiff,
UK. You will need to use WinMDI to prepare files for analysis.
Cytometry Shareware
FCS Press and FCS Assistant
This software will you let you edit FCS files and create presentation
quality graphics. This software is provided by Ray Hicks and costs about $50,
plus $5 for each additional licence.