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FACSCalibur

The FACSCalibur has 4 fluorescence detectors plus forward- and side-scatter detectors.  See the table below to determine which fluorochromes may be used. 

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 software to collect and analyze data.  Using CellQuest, data may be analyzed on dot plots and histograms with various gates.

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

The FACSCalibur is connected to the UAMS network.  Data from the Calibur may be stored temporarily on the network at \\resnet\FACS in a designated folder.  Otherwise, data may be stored on CD or Zip disk directly from the FACSCalibur computer.

 

 

 

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