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FacsCalibur

 

The FACSCalibur has 4 fluorescence detectors plus forward- and side-scatter detectors. The FACSCalibur is used for analysis only. The Calibur can analyze up to four colors at one time. The Calibur is also the instrument used for cell cycle, cell proliferation, cell membrane potential, and other analysis. To determine if your fluorochromes can be analyzed on the FACSCalibur, see the table below to determine which fluorochromes may be used based on your fluorochromes excitation and emission. 

 

Keep in mind that many fluorochromes will have overlapping emission spectra, and may be difficult to distinguish when used together.  Be sure to check the excitation and emission spectra of the fluorochromes you plan to use to optimize your experiment.  The BD Biosciences Fluorescence Spectrum Viewer may help you determine whether the fluorochromes you want to use will have a significant overlap.  We also have a list of some fluorochromes, dyes and fluorescent proteins you may check. 

Excitation Laser

Detection Filter

Detector

Example

488 nm (blue)  

488/10 (483-493 nm)

FSC Diode

FSC

 

488/10 (483-493 nm)

SSC PMT

SSC

 

530/30 (515-545 nm)

FL-1 PMT

FITC

 

585/42 (564-606 nm)

FL-2 PMT

PE

 

670LP (>/= 670 nm)

FL-3 PMT

PerCP

635 nm (red)

661/16 (653-669 nm)

FL-4 PMT

APC

Analysis

The FACSCalibur computer uses CellQuest Pro software to collect and analyze data.  Your data will be analyzed as part of your flow cytometry appointment as long as time allows, or you may set an appointment specifically for data analysis.  Because analysis requires time on the cytometer computer, analysis-only appointments are charged the same rate as data acquisition/analysis.  You may choose to work with facility personnel to analyze your data, or you may analyze data on your own. 

WinMDI is a free program available for use on PC's.  There is also a tutorial for the program to help you learn to use it.  Other programs are also available under our useful links menu. 

Data Delivery

Please bring a USB drive or blank CD to save your data files. The Core Facility does not store or archive data for longer than about 6 months so it is your responsibility to get the data files and save them for future use.

 

 

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