Flow Cytometry Shared Resource

On this page, you will find:

About Us
Areas of Focus
Services We Provide
Directors
Published Research
Contact Us

About Us

The Flow Cytometry Shared Resource provides researchers with ready access to the necessary expertise and state-of-the-art instrumentation they need to explore a wide range of research problems with flow cytometry.

We focus on addressing the needs of seasoned flow cytometry users, as well as those just discovering the capabilities of flow cytometry technology. We provide monthly training for the instruments for new users. The directors are also available to consult on experimental design and data analysis.

Areas of Focus

The typical experiments handled by the Flow Cytometry Shared Resource Facility are designed to do analysis or to sort based on up to 9 fluorescent parameters. Some representative examples of analysis experiments include the:

Measurement of DNA and RNA content for cell cycle analysis
Cell cycle specific nuclear and cytoplasmic proteins
Apoptosis markers
Substituted deoxyuridine incorporation
Stem cell side populations
Reactive oxygen species
Cell surface markers using labeled antibodies or substrates
Cell viability
Intracellular ion concentration or pH
Membrane potential

Cell lysates can also be analyzed with bead substrate based immunoassays. Some representative examples of sorting experiments include:

Purification of cells transfected with fluorescent protein tagged DNA inserts
Isolation of cell populations expressing specific intracellular or membrane metabolic products.

Services We Provide

Services we provide include:

Use of the Instruments
Consultative Services
Training

Use of the Instruments

The institutional instruments available as part of the Flow Cytometry Shared Resource include:

BD FACS Aria cell sorter (3 lasers and 11 parameters)
- Sorting tips available: 70 micron, 100 micron & 130 micron.
- Instrument is fitted with a special flow cell to provide better precision for measuring DNA content and side-populations
BD FACS Canto II analyzer (3 lasers and 10 parameters)*
Two BD FACS Calibur analyzers (2 lasers and 6 parameters)*
In addition to the software available with each flow cytometer, FCS Express flow cytometer analysis software is available to analyze the multiparameter data to identify and characterize cell subpopulations. DNA histogram analysis software programs (ModFit and MultiCycle) are also available to address the needs of researchers requiring advanced cell cycle analysis.

* Instruments are available to use 24 hours, 7 days a week. Users must seek the appropriate instruction to be approved instrument operators. See below.

Reservations to Use the Equipment

Investigators reserve time on the FACS Caliburs and FACS Canto via a Web-based public calendar that is accessible to all institutional users.

Reservations for sorting time on the FACS Aria are made by direct communication with Dr. James Wood or Beth Holbrook, as these individuals are responsible for performing the sorts. (See contact information below). Cost for instrument utilization is as follows:

FACSCalibur: $45/hour
FACSCanto: $50/hour
FACS Aria: cell sorting $50/hour plus a $25 set-up fee

Cancer Center members performing cancer related studies receive a 50 percent subsidy.

Consultative Services

The directors provide consultative services at no charge to the investigator for discussions regarding experiment design, execution or interpretation. In many cases, this saves the investigator time and resources by preventing mistakes in the initial experimental design that may not be obvious to less experienced users. Investigators also have access to the wide network of pre-eminent flow cytometry users from academia and industry via personal introductions by Dr. Wood, who has extensive connections in this field.

Training

Hands-on training is provided for individuals who would like the ability to acquire their data on either the FACS Caliburs or FACS Canto. Dr. Martha Alexander-Miller should be contacted directly to schedule for the one of the monthly training sessions. (See contact information below)

Directors

Martha Alexander-Miller, PhD, Director
James C. S. Wood, PhD, Manager

Published Research

Seamer L, Bagwell CB, Barden L, Redelman D, Salzman G, Wood JCS, Murphy RF: “Proposed New Data File Standard for Flow Cytometry, Version FCS 3.0.” Cytometry. 28:118-122, 1997.
Wood JCS, Hoffman RA: “Evaluating Fluorescence Sensitivity on Flow Cytometers: An Overview.” Cytometry. 33: 256-259, 1998.
Wood JCS: “Fundamental Flow Cytometer Properties Governing Sensitivity and Resolution.” Cytometry. 33:260-266, 1998.
Novo D, Wood J. “Flow cytometry histograms: transformations, resolution, and display. Cytometry Part A. 73A:685-692. 2008.
Lee JA, Spidlen J, Boyce K, Cai J, Crosbie N, Dalphin M, Furlong J, Gasparetto M, Goldberg M, Goralczyk EM, Hyun B, Jansen K, Kollmann T, Kong L, Leif R, McWeeney S, Moloshok T, Moore W, Nolan G, Nolan J, Nikolich-Zugich J, Parrish D, Purcell B, Qian Y, Selvaraj B, Smith C, Tchuvatkina O, Wertheimer A, Wilkinson P, Wilson C, Wood J, Zigon R, The International Society for Advancement of Cytometry Data Standards Task
Force, Scheuermann RH, Brinkman RR “MIFlowCyt: The minimum information about a flow cytometry experiment.” Cytometry Part A. 73A: 926-930. 2008.
Spidlen J, Leif RC, Moore W, Roederer M, International Society for the Advancement of Cytometry Data Standards Task Force, Brinkman, R: “Gating-ML: XML-based gating descriptions in flow cytometry.” Cytometry Part A. 73A:1151-1157. 2008.
Spidlen J, Moore W, Parks D, Goldberg M, Bray C, Bierre P, Gorombey P, Hyun B, Hubbard M, Lange S, Lefebvre R, Leif R, Novo D, Ostruszka L, Treister A, Wood J, Murphy RF, Roederer M, Sudar D, Zigon R, Brinkman RR: “Data File Standard for Flow Cytometry, version FCS 3.1.” Cytometry Part A. 2010; 77 (1): 97-100.
Wood JCS: “Clinical Flow Cytometry Instrumentation.” In Clinical Flow Cytometry: Principles and Application (K. D. Bauer, R. E. Duque, T. V. Shankey, eds.) Williams and Wilkins, Baltimore, 1993. pp 71-92.
Wood JCS: “Unit 1.8. Principles of Gating.” In Current Protocols in Cytometry (J. P. Robinson, Z. Darzynkiewicz, P. N. Dean, A. Orfao, P. S. Rabinovitch, C. C. Stewart, H. J. Tanke, eds.) John Wiley & Sons, Inc., New York, 1999.
Wood JCS, Hoffman RA: “Unit 1.20. Characterization of Flow Cytometer Instrument Sensitivity.” In Current Protocols in Cytometry (J. P. Robinson, Z. Darzynkiewicz, P. N. Dean, A. Orfao, P. S. Rabinovitch, C. C. Stewart, H. J. Tanke, eds.) John Wiley & Sons, Inc., New York. 2007.
Wood JCS: “Unit 1.4. Establishing and Maintaining System Linearity.” In Current Protocols in Cytometry (J. P. Robinson, Z. Darzynkiewicz, P. N. Dean, A. Orfao, P. S. Rabinovitch, C. C. Stewart, H. J. Tanke, eds.) John Wiley & Sons, Inc., New York, Revised 2009.