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College of Veterinary Medicine

Diagnostic Medicine/Pathobiology

Flow Cytometry Lab

The Flow Cytometry laboratory is housed in Mosier Hall, Room K208 of the College of Veterinary Medicine Complex. This multi-user core laboratory is equipped with state of the art flow cytometry analyzers by Becton Dickinson (LSR Fortessa X-20 and FACSCalibur), a multiplex MagPix instrument, and fluorescence activated cell sorting equipment by BioRad, an S3 air-in-stream cell sorter within a BSL-2 hood.

Hours of Operation: M – F 8 – 5 pm, Call to schedule a time on the instruments

Usage Fees
InstrumentInternalExternal
BD FACSCalibur$20/15min$30/15min
LSR Fortessa X20$25/15min$37.50/15min
Assisted flow on Analyzers$30/15min 
BioRad S3 Cell Sorter$45/15min$67.50/15min
MagPix Setup (by staff)$30/plate$45/plate
TRAINING PACKAGES (2 hrs)$100 

Subsidized by Department of Diagnostic Medicine/Pathobiology/KSVDL.

New users will be strongly encouraged to sign up for a minimum of 2 hours of training. Training will cost $100, and will accommodate up to 2 trainees per session. The training sessions will cover instrument set-up, data acquisition, trouble-shooting, and introduction to analysis software.

E-mail or call for consultation
Director

Melinda J. Wilkerson, DVM PhD
wilkersn@vet.k-state.edu
Ph: 785-532-4818

Research Associate

Kaori Knights, MS
kknights@vet.k-state.edu
Ph: 785-532-4617
Fax: 785-532-4072

Research Assistant (Flow Cytometry certified operator)

Anushka George, MS
ageorge@vet.k-state.edu
Ph: 785-532-4617
Fax: 785-532-4072

 

Instrument

Lasers

Fluorescent Markers

FACS CALIBUR488nm, 642nmFITC, GFP, Alexa 488, PE, PI, PerCP, 7-AAD, Pe-Cy7, APC, Alexa 647
BioRad S3 Cell Sorter488 nm, 640 nmFITC, GFP, Alexa 488, PE, PI, PETxRed, APC, Alexa 647, PerCP, PE-Cy7, APC-Cy7, Alexa 700, or any equivalent wavelength fluorophores.

LSR Fortessa X-20 Special Order Research Product Flow Cytometer

405nm, 488nm, 561nm, 640nmFITC, GFP, Alexa 488, Alexa 546, Alexa 647, PerCP-Cy5.5, APC, Alexa 700, APC-Cy7, PE, PI, 7-AAD, PerCP, PETxRed, PE-Cy5, PE-Cy7, BV421, PacBlue, BV510, AmCyan, BV650, BV785, or any equivalent wavelength fluorophores.

 

FACSCalibur

The FACSCalibur is equipped with two lasers that emit light at 488 nm and 630 nm. There are three detectors (FL1, FL2, FL3) for the 488 laser and one detector (FL4) for the 630 laser.  This two laser cytometer allows the operator to analyze up to 4-color and 2 size parameters of cells or fluorescent-labeled particles. FACSCalibur is interfaced with a Macintosh computer and a publication quality color printer. Cell Quest and ModFit analysis software is available on two computers to analyze flow cytometry data including DNA and proliferation assays.

BioRad S3 Cell Sorter

The BioRad S3 is a stream-in-air cell sorter equipped with two co-linear lasers (488 and 633 nm) having 4-color detection capabilities. This instrument can determine size, complexity, and 2-way sort cells.  The sorting operation is fully automated. This machine was previously equipped with a 561 yellow-green laser which has now been replaced with the 633 red laser. The upgrade has altered the fluorophores detected by the sorter; please check the fluorescent marker chart for current compatibility. This sorter accepts samples in 5.0 mL falcon tube format and can sort into1.5 mL eppendorf tubes or 5.0 mL falcon tubes. Polypropylene tubes are highly recommended for all sorting applications in lieu of the standard polystyrene tubes. The sorter is only operated by core facility staff. Please contact the staff directly before signing up to use the cell sorter.

LSR Fortessa X-20 Special Order Research Product Flow Cytometer

The Fortessa X-20 is equipped with 4 non-linear lasers (405, 488, 561, 640 nm) and filter configurations allowing for detection of up to 12 colors plus Fwd and Ssc parameters. The cytometer utilizes FACSDiva v8.0 acquisition software and generates FCS files compatible with all commercial analysis programs. Routine CS&T calibration allows for standardization between experiments, increasing the reliability of data generated over time. This cytometer accepts samples in 5.0 mL falcon tube format with a recommended sample volume of approx. 300+ uL.

MagPix

The MagPix by Millipore can analyze up to 50 analytes in a single sample. Examples are cytokine, chemokine, and serological panels. Call for consultation and quote for usage.

Additional educational resource: Tutorials on flow cytometry