All users receive basic education on the science and technology of flow cytometry, cell sorting, and immunoassays through both formal and informal programs. Bi-monthly formal lectures cover the basic concepts of flow cytometry, including compensation, fluorochrome selection for multi-color assays, sample preparation, software for data analysis, together with the principles of each immunoassay that is available for use. This lecture series occurs approximately three times per year. These lectures are posted on our website, together with self-help modules.
Errors in sample acquisition and/or data analysis are common among poorly trained users and can lead to misleading and/or unreliable data. We have a room in the Flow Cytometry Shared Resource that is dedicated to investigator-performed data analysis. There are 5 work-stations, including 3 G4 MacPro's, a 27" iMAC and a G5 PowerPC with dual 27" monitors. Flow cytometry software includes Cell Quest, FlowJo, Modfit, and Winlist. The Shared Resource staff is on hand to field questions. We are beginning to phase in GemStone (Verity Software House, Topsham, ME) software, a new paradigm for analyzing high-dimensional, flow cytometry data based on probability state modeling. We have also introduced Omniviz (BioWisdom Ltd, Cambridge, UK) software to simplify analysis of mechanistic studies that have multiple patients, each with multiple time points.
- Introduction to flow cytometry
- Compensation and FMOs
- Lasers, filters and fluorochromes
- Optimizing fluorochromes for multi-color flow cytometry
- Experimental design
- Cell sorting
- Data analysis
- Flow cytometry software (FlowJo, Modfit, Winlist, FCSExpress, Kaluza, Gemstone, Spice, etc.)
- Multiplex cytokine assays
Instrument training is available for individual researchers by an experienced staff member and covers basic operational procedures including acquisition software and Shared Resource policies. Initial training at the flow cytometry instrument involves standard samples. Subsequent training sessions involve a user’s own samples – with training specific to the needs of that user. At following sessions, one-on-one supervision is provided for as long as required, with the eventual goal of making the scientist, student, or technician a careful and independent user of the facility.
A critical component to this training is the effective use of written, up-to-date standard operating procedures (SOPs), which are available on this website as well as at each instrument. Special applications (intracellular cytokines, cell proliferation, apoptosis, phosphorylation assays, etc) are offered as lecture/wet laboratory formats in a Flow Cytometry Symposium each year. In addition, we invite outside speakers to the Department of Immunology and Microbiology seminar series to discuss advanced technology.
We actively encourage investigator-performed services, and the shared resource is accessible 24 hours a day, 7 days a week. Those users who choose to run experiments after hours when Shared Resource staff is not present, are trained to a high standard, like the training received by our technicians. This includes instruction on instrument startup operations, daily QC procedures, acquisition techniques, basic cleaning operations, and shutdown operations. Although cell sorting requires a much higher level of expertise, and much more training time, our policy is to train high-frequency users so they can use the instrument after hours. In this way, the Shared Resource can maximally utilize the instruments, increase usage, while controlling total staffing costs.
The training effort is a significant time investment, but the overall benefit is both cost-effective and promotes higher quality data and supports the educational mission of an academic research center.