1.  What is the Biological Hypothesis?

2.  What cell subsets need to be identified?

              Primary Importance?

              Secondary Importance?

              Tertiary Importance?

              Luxury

3.  Choose antigens for identifying subsets:

4.  What fluorochromes are available?

              Which instrument are you using?

                            Lasers?

                            Fluorochromes the instrument can detect?

              What conjugates are available for each antigen?

5.  Design staining combinations:

              High Density Antigens-

              Medium Density Antigens-

              Low Density Antigens-

6.  Is a viable dye required?

7.  Is fixation required?

8.  Controls:

              Compensation-

              FMO-

              Experimental-

9.  Draw out Analysis Plots/gating strategy. What statistics are required?

10. Is there a time requirement for analysis following staining?