Services
The Core offers many services for the analysis or isolation of cells following staining of cellular proteins or DNA with fluorochrome labeled reagents.
Some of the FACS applications include:
- Immunophenotyping
- Intracellular staining
- Molecular and cellular probes (Ca flux, GFP, DNA cell cycle)
- Analysis of cells undergoing cell death or apoptosis (TUNEL assay, Annexin V)
- Detection of soluble proteins via bead capture (Cytometric Bead Array, Flowcytomix)
- Sterile isolation of selected cell populations on the basis of specific protein expression or cell size
Data is acquired as FCS 3.0 using BD Cell Quest V3.3 on the FACScan and FACSVantage. Additionally, BD FACSDiva Software V5.0 is used to acquire data on the LSRII. Data can be imported/exported as FCS 2.0 or 3.0 when using FACSDiva. List mode acquisition and storage of data allows for analysis and presentation many different formats according to the needs of the investigator. Data files are permanently stored in personal folders on the LRI servers, readily available to all LRI employees with proper logon accounts. CellQuest, FlowJo and ModFit software are provided for analysis. Investigators are encouraged to schedule a training session to learn analysis of flow data utilizing one of the software packages available.
Consultation
All investigators are encouraged to meet with the Flow Cytometry Core technologists to discuss the experiment. This meeting is often useful in that the technical limitations of an experiment are realized and many potential problems can be averted before critical resources are used. The Core can also provide written protocols for staining, fixation and data analysis methods. The Core technologists are available for assistance with data retrieval, analysis and presentation.
Materials available for reference in the Flow Core lab include several textbooks on the basics of flow cytometry and the Current Protocols in Cytometry. In addition, the Core frequently organizes training seminars at different levels of expertise.